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Abbreviations

Reactivity:

Bab: Baboon; Bov: Bovine; C: Chicken; Cyno: Cynomolgus; D: Dog; Dros: Drosophila; F: Frog; G Pig: Guinea Pig; Gt: Goat; Ham: Hamster; Hs: Horse; Hu: Human; Koa: Koala; Mam: Mammalian; Ms: Mouse; Pig: Pigeon; Qua: Quail; R: Rat; Rb: Rabbit; Rhe: Rhesus; Tr: Trout; VTC: Viral transformed cells; Xen: Xenopus

Applications:

Block: Blocking; FA: Functional Assay; FCM: Flow Cytometry; IC/FCM: Intracellular Flow Cytometry; IF: Immunofluorescence; IHC: Immunohistochemistry; IHC(Fr): Frozen; IHC(F): Formalin; IHC(P): Paraffin; IHC(Zn): Zinc; IP: Immunoprecipitation; Neu: Neutralizing; RIA: Radioimmunoassay; RPA: Ribonuclease Protection Assay; WB: Western blot

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Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5Review Article on the Field of Neuroscience . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Advanced Culture Surfaces for Neuronal Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

BD Falcon Cultureware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

BD Falcon Pipets and Tubes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

BD Primaria Cultureware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

Extracellular Matrix Proteins and Attachment Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

BD Matrigel Basement Membrane Matrix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

BD BioCoat Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

BD BioCoat Matrigel™ Matrix Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

BD BioCoat Laminin Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

BD BioCoat Collagen IV Cellware. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

BD BioCoat Poly-Lysine Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

BD BioCoat Poly-D-Lysine/Laminin, Poly-L-Orinthine/Laminin, and Laminin/Fibronectin Cellware . . . . . . . . . . . . . . . . . 25

BD BioCoat Collagen I Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

BD BioCoat Fibronectin Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

BD BioCoat Gelatin Cellware . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Cell Culture Inserts and Insert Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

BD BioCoat Services . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

BD Growth Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Functional Assays and Discovery Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32Apoptosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Cell Signaling, including Kinases and Phosphotases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Spotlight Phosphoprotein Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

BD Phosflow Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Cytoskeletal Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

Development and Differentiation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

Growth Factor Related . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

Ion Channels and Transporters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48

Neurotransmitter Receptor-Related . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Synaptic Vesicles and Post-Synaptic Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51

BD Assay Kits and Reagents for Bioimaging and Flow Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54

BD Cytometric Bead Array Flex Set Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55

Technical Spotlight and Protocols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60Screening and Development of Antibodies for use in Cell-Based Assays using Immunofluorescence Microscopy . . . . . . . . . 60

Phospho-Protein Profiling in Stem Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63

IHC for Neuroscience Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64

Phospho-Specific Staining of Rat Brain Tissues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

Imaging and High Content Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66Bioimaging Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66

Cellular Analysis Using Flow Cytometry. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

BD Flow Cytometry Instrumentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70

bdbiosciences.com3Unless otherwise specified, all products are for Research Use Only. Not intended for use in diagnostic or therapeutic procedures. Not for resale.


Table of Contents



BD Biosciences Neuroscience Source

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releases, and technical updates.



Introduction

Neuroscience is a rapidly changing and expanding area of research with the goal of

deciphering the brain’s neurological systems and related disorders, such as

Parkinson’s and Alzheimer’s diseases. BD Biosciences is proud to be an integral participant

in this important research.

Imagingand

High ContentAnalysis

Cell Culture

Functional Assaysand Discovery Reagents



Review Article on the Field of Neuroscience

Benjamin Wolozin, MD, PhDProfessor of PharmacologyDepartment of Pharmacology and Experimental TherapeuticsBoston University School of MedicineBoston, MA, USA

A decade of research has led to dramatic

advances in our understanding of the

causes of many diseases of the central

nervous system. One of the striking

observations is the presence of common

biochemical pathways that contribute to

the pathophysiology of a wide variety of

diseases. Neurons, glia and oligodendro-

cytes subjected to varying types of stress

or injury show activation of many common

pathways such as the stress kinase cascades

and the apoptotic cascades. Identification

of these common signaling pathways

provides researchers with the biochemical

framework to explore disease pathways

for known illnesses and new genetic vari-

ants of illnesses. In some cases the central

pathways of disease are known, but the

small tributaries that subtly differentiate

diseases, particularly late onset diseases,

are not understood. Many of these

signaling processes often occur in parallel.

At the same time, fundamental questions

remain. The subtle reasons why one person

develops Alzheimer’s disease, a second

person develops Parkinson’s disease, and

a third is still healthy at 95 years of age

remains a mystery.

Proteins causing familial neurodegenerative diseases

Common diseases of the central nervous

system fall into three general categories:

late onset neurodegenerative diseases, acute

injury, and psychiatric illnesses. The response

of the brain to neurodegenerative diseases

such as Alzheimer’s disease, Parkinson’s

disease, Huntington’s disease and amyo-

trophic lateral sclerosis is pleitropic. Familial

forms of each of these diseases are associ-

ated with abnormalities in a specific protein.

Alzheimer’s disease: Alzheimer’s disease

(AD) is characterized by the accumulation

of neuritic plaques containing aggregated

b-amyloid (Ab) and neurofibrillary tangles

containing fibrillar microtubule associated

protein tau. In AD, mutations in genes that

regulate production of Ab lead to early

onset familial AD. Mutations in amyloid

precursor protein, presenilin 1 and presenilin

2 all cause familial AD. Discovery of these

genes also lead to discovery of the b and

g-secretase complexes. b-Secretase activity

is largely attributed to the action of one

membrane associated protein, BACE 1.

g-Secretase is composed of at least four

different membrane associated proteins:

presenilin, nicastrin, pen-2 and aph-1.

Increased production of total Ab or Ab42,

a longer form of Ab that aggregates

rapidly, leads to increased deposition and

causes neurotoxicity. This neurotoxicity is

accompanied by tau hyper-phosphorylation

and formation of neurofibrillary tangles

composed of tau protein. Proper functioning

of tau is clearly important for neuronal

health because mutations in tau are

associated with a different type of dementia,

termed fronto-temporal dementia.

One of the challenges facing researchers

investigating AD is the distinction between

the uncommon forms of early onset, familial

AD that are associated with mutations in

presenilin or APP, and the more common

forms of disease that tend to occur later

in life. The apolipoprotein E4 isoform is

the only polymorphism clearly associated

with late onset AD. Polymorphisms in other

proteins, such as the ubiquilin 1 protein,

might be associated with late onset AD, but

the associations remain controversial and

the strength of the effect is far below that

of apolipoprotein E4.

Parkinson’s disease: At least 5 different

genes have been associated with familial

Parkinson’s disease (PD). a-Synuclein was

the first protein linked to familial PD.

Understanding the biology of a-synuclein

attracted extensive attention because it

is the main protein that accumulates in

Lewy bodies, which are the pathological

hallmark of sporadic PD. The involvement of

a-synuclein in both familial PD and sporadic

PD provides a critical linkage between

familial PD and sporadic PD. Research into

the pathophysiology of a-synuclein indicates

that this protein tends to aggregate, and

that the mutations associated with familial

disease increase this tendency to aggregate.

The tendency of a-synuclein to aggregate

presents a common theme that is apparent

in many neurodegenerative diseases

because many of the pathogenic proteins

tend to aggregate. Aggregation, or more

specifically, oligomerization and fibrillization,

are essential steps in toxicity related to

both Ab and tau toxicity. Four other genes

have been clearly associated with familial

Parkinson’s disease, and more genes carrying

mutations associated with familial PD are

likely to be identified. Loss of functions

mutations in parkin, DJ-1 and Pink-1 all

cause familial PD, while mutations in LRRK2

associated with familial PD are thought to

cause a gain of function. Parkin is an E3-

ubiquitin ligase; LRRK2 binds parkin. The

role of E3-ligases in proteasomal function

suggests that proteasomal dysfunction

could contribute to familial PD. On the other

hand, toxicological studies indicate that

chemicals inhibiting mitochondrial function,

such as MPTP or rotenone, can mimic

some aspects of PD. Pink-1, DJ-1, LRRK2

and parkin all associate with mitochondrial

structures, and neurodegeneration caused

by over-expressing a-synuclein also

causes mitochondrial degeneration. These

studies present tantalizing clues about

the causes of PD, but a comprehensive

understanding of the mechanism of

degeneration remains elusive.

Disorders of the Nervous System: New Tools to Examine New Pathways



Polyglutamine-associated neurodegenerative disorders: Expansions of polyglutamine-repeat

domains are associated with 7 different

diseases, including Huntington’s disease,

Spinobulbar muscular atrophy, 6 forms of

spinocerebellar ataxias and dentatorubral-

palidoluysian atrophy. Proteins harboring

expanded polyglutamine repeats aggregate

readily. The aggregates accumulate in the

nucleus or the cytoplasm, depending on the

cell type. Aggregation in the cytoplasm or

endoplasmic reticulum causes proteasomal

inhibition. Nuclear aggregation is associated

with transcriptional changes, possibly

because some transcription factors, such

as CREB, contain polyglutamine repeat

domains and tend to be sequestered by the

nuclear aggregates. Studies using animal

models suggest that loss of CREB function in

polyglutamine repeat disorders contributes

to the neuronal dysfunction. Treatment

with HDAC inhibitors counteracts the loss

of CREB function and is now being tested

in patients with Huntington’s disease.1

Amyotrophic Lateral Sclerosis: The

mechanisms producing amyotrophic lateral

sclerosis (ALS) remain enigmatic. ALS is an

uncommon disease. Familial ALS, which

accounts for about 10% of the cases,

is caused by mutations in superoxide

dismutase. Although the mutations increase

the tendency of superoxide dismutase

to aggregate, the mutations appear to

cause disease by altering the activity of

superoxide dismutase, perhaps by causing

release of the copper or zinc at its core,

rather than by a mechanism linked to

aggregation. The relationship between

familial and sporadic ALS, though, remains

elusive. Sporadic ALS might be linked

to dysfunction of glutamate uptake by

glia, but this theory is controversial.

Different diseases converging onto common pathways

Understanding why each particular

biochemical abnormality is deleterious to

the brain represents one of the central goals

of research for many of these diseases. The

biochemical dysfunction that underlies each

disease induces a number of processes that

are detrimental to the cell including produc-

tion of reactive oxygen species, activation

of stress kinases and ultimately activa-

tion of the apoptotic signaling cascade.

These processes are also stimulated during

the types of acute injury that occur with

trauma, stroke, status epilepticus and

binging on addictive drugs. Despite a wide

variety of different insults that initiate the

neuronal injury, the brain shows cellular

responses that generalize across a broad

number of diseases.

The stress kinase cascade flows through

a large number of MAP kinases and

MAPKAP kinases to converge on a number

of terminal kinases that include the c-Jun

kinase and p38 family of stress kinases.

DNA damage activates repair systems

through enzyme cascades mediated by p53

and other checkpoint proteins. The p53

protein also regulates a large number of

cellular and mitochondrial responses. Small

gaseous signaling agents, such as nitric

oxide and carbon monoxide, modulate

signaling cascades that can be good or bad,

depending on the cell type. The apoptotic

cascade flows to and then emanates from

the mitochondria. This cascade proceeds

via a number of linked biochemical reac-

tions including caspase activation, reaction

by the Bcl-2 family of proteins, release of

cytochrome C, and modulation of other

proteins that regulate apoptosis such as IAP

and Diablo. The mitochondria respond to

the damage, producing free radicals due

to electron leakage and reverse electron

transport. The production of free radicals

combined with changes in calcium content

and other signals act to open the perme-

ability transition pore. Increased calcium

also causes damage by activating calcium

dependent proteases, and the energy

demands that are sometimes associated

with excitotoxicity put further strain on the

capacity of the mitochondria, driving down

cellular energy stores.

Extrinsic signaling also modifies neurode-

generation. Degeneration in the CNS is

frequently accompanied by an activated

inflammatory response, which is presum-

ably designed to remove the damaged

tissues. Inappropriate activation of immune

responses in the brain lies at the heart of

multiple sclerosis. The cytokines and reac-

tive oxygen species (ROS) produced by the

microglia are directly toxic to oligodendro-

cytes and neurons; production of cytokines

and ROS plays an important role acceler-

ating degeneration in AD, PD and ALS.

Understanding how to dampen or direct

the immune response represents one of

the many challenges facing researchers of

neurodegeneration.

The degenerative processes can be counter-

acted by growth factors, which activate

signal transduction cascades that promote

survival and cell growth. These cascades

induce a reactive response that stimulates

synaptic plasticity at the same time as other

elements of the neurons are undergoing

degeneration. Many studies have shown

that factors that stimulate pro-survival

cascades delay degeneration. Brain derived

nerve growth factor, nerve growth factor

and glial derived growth factor all have

been shown to be neuroprotective in

cellular and animal models.

The brain also has the capacity to respond

to the neuronal injury by producing new

neurons in some brain areas from stem

cells. These pluripotent cells can replace lost

neurons, and ultimately might represent an

endogenous source of cellular replenish-

ment for the brain. The cytokines and ROS

produced by the disease process, though,

appear to harm stem cell production in

addition to inducing neurodegeneration.

Many studies show that the capacity to

produce stem cells is reduced in animal

models of neurodegenerative disease.



Review Article on the Field of Neuroscience (continued)

Each of these factors – signaling, growth

factors and regeneration – impinge on

the neuron to modulate synaptic function.

Increases in our understanding of synaptic

biology have lead to dramatic advances

in our understanding of the biochemical

mechanisms underlying diseases such as

schizophrenia, depression, drug addic-

tion and autism. The synapse is a finely

tuned microscopic engine not dissimilar

to a nano-robot. It has numerous inter-

connected elements that must respond

in concert to achieve the optimal level

of neuronal signaling. Subtle changes in

the amount or type of signaling lead to

dramatic changes in disorders that manifest

in altered behavior. Families of proteins,

such as neuregulins and dysbindin, modu-

late activity of the glutamate receptors.

Mutations at different points in this regula-

tory cascade appear able to predispose to

schizophrenia when environmental insults

are also present to stress the system. For

instance, mutations in neuregulin I type 3

cause dysfunction of glutamate receptors

and cholinergic neurons and appear to be

linked to schizophrenia.

Serotonin neurotransmission plays a central

role in depression, with low serotonin

precipitating depression. Medications, such

as the selective serotonin reuptake inhibi-

tors (SSRIs), can alleviate the depression

but the slow kinetics of the response of

depressed patients to SSRIs suggests that

other processes need to occur in parallel.

SSRIs are known to stimulate growth factor

responses and also have been recently

shown to slowly modulate a protein termed

P10. These processes provide important

clues about the pathophysiology of depres-

sion. Activity of dopaminergic neurons is

associated with drug abuse, but too much

activity stimulates glutatamate-mediated

neurotoxicity. In addition, the overwhelming

pleasure stimulated by drugs of abuse and

the resulting craving produce incapacitating

behavioral responses. Medications that

reduce the strength of the response, such

as naltrexone, show promise for treatment

of some addictions, but understanding how

to intervene in this circuitry remains a major

challenge in the field of behavioral neurosci-

ence. Our increasing understanding of the

cell biology of the addictive axis provides

a corresponding increase in the number of

strategies and targets for intervention.

High throughput technologies: Arrays, multiplexing and systems biology

Disease processes are typically complex.

New technology allows the investigator to

characterize multiple responses occurring

in parallel. High throughput technology

has emerged as a key approach towards

dissecting the complex series of biochemical

events that occur in parallel in the brain.

This technology can provide a broad over-

view of the magnitude and kinetics of

changes in gene expression, protein expres-

sion or post-translational modifications.

Gene arrays provide an overview of the

broad spectrum of transcriptional responses

to any particular signal. Three different

technologies are also emerging to provide

overviews of the response of the proteome

in stressed or injured tissues. 1) Antibody

arrays allow analysis of how the levels of

hundreds or thousands of proteins respond

to a stress, such as treatment with Ab. 2)

Two-dimensional gel technology combined

with mass spectroscopy also allows identifi-

cation of proteins that might not be picked

up through antibody arrays. 3) Multiplexing

provides accurate quantification of dozens

of targets in one rapid assay.

Multiplexing is the term used to describe

assays that measure multiple epitopes

at once. Bead technology allows parallel

analyses of multiple epitopes in one assay

using either dedicated multiplexing devices

or BD FACS analyzers, which have the

benefit of being available to many labora-

tories through institutional core facilities.

Multiplexing allows any laboratory to

affordably profile metabolic responses of

tissues. Multiplexing technology is also

highly valuable for situations where the

tissues are available in only limited

quantities. By analyzing multiple proteins in

parallel in the same sample, researchers are

able to maximize the information gained

from small amounts of sample. The ability

to dissect, categorize and quantify protein

networks in tissues is increasingly being

applied to facilitate translational research

in an effort to move today’s breakthroughs

into tomorrow’s medicines.

Reagents are now available to identify and

quantify the cascades of protein activity

associated with stimulation of biochemical

pathways. Antibodies specific to phosphory-

lated proteins allow determination of the

amount of activation of a particular protein

and the kinetics of activation. Patterns of

activity of proteins can be studied for indi-

vidual proteins or for groups of proteins.

The latter analysis allows comparison of

activities of different pathways or validation

of activation by analysis of multiple steps

in a particular signal transduction pathway.

The commercial sector offers strong libraries

of reagents to facilitate studies of signal

transduction.

Imaging: The science of real estate

Accumulating evidence demonstrates the

immense importance of location in neuro-

biology. The delineation of whether an

inclusion occurs in the nucleus, cytoplasm,

mitochondria or extracellular domain has

important implications for mechanisms

of degeneration and potential avenues of

therapy. Identifying which synapses are

subject to oxidative damage or post-trans-

lational modification focuses attention on

particular neuronal cell types and particular

receptor interactions. Determining whether

inflammation targets a neuron, a glia or

an oligodendrocyte helps to define the

disease process. The steady march of cell

culture methods facilitates our knowledge

of neurobiology by allowing researchers

to analyze isolated groups of cells, but

novel imaging technologies, novel antibody

probes and novel fluorophores greatly

expand our imaging capabilities. Multiple

excellent fluorophores are now available to



facilitate double, triple and even quadruple

staining protocols. Confocal microscopy

allows 3-dimensional reconstructions of

molecular distributions within a neuron or

glial cells. Using confocal microscopy we

can determine whether epitopes are actually

co-localized or simply vertically juxtaposed.

Imaging technologies such as fluorescence

resonance transfer can test whether puta-

tive molecular interactions actually occur

in living cells. Imaging has also moved in

vivo. Magnetic resonance imaging and

positron emission tomography has been

used for years to probe the brain, but now

techniques such as two-photon imaging

allows analysis of cellular events in the living

brain. Developing imaging technology and

improved biomarkers offers the ability to

study structures such as amyloid plaques

and neurofibrillary tangles in living animals.

The future of disease research is likely

to shift towards more complex analyses

as systems biology is applied to disease

processes. Massive parallel studies of gene

arrays and protein interaction screens are

already revolutionizing our understanding

of the genetic events that take place in

a developing nematode or proliferating

bacteria. This type of technology will

increasingly be applied to understand the

human body, both in health and in disease.

Multiplexing will need to be applied to

bridge the gap between gene arrays,

systems biology and disease research.

Identifying key signaling pathways and

quantifying the selective changes that lead

to particular diseases will certainly facilitate

developing diagnostic tests and therapeutic

treatments for the diseases that plague our

populations today.



Cell Culture

Applying Innovative Technology to Neuronal Cell Culture

As the first company to produce sterile,

disposable labware more than 50 years

ago, BD Biosciences is a worldwide leader

in top-quality cell culture products. Our

comprehensive line of innovative products

will meet your neuronal cell culture needs.

Consistent cell culture conditions are

required for reproducible research results.

For many cell lines, the easiest solution is

to use a treated surface that allows cells

to attach. The consistency of this surface

depends on the treatment method used.

Many manufacturers have long used atmos-

pheric plasma treatments (eg, corona) to

create hydrophilic surfaces. In corona treat-

ment, a high-voltage discharge creates

a reactive gas plasma above the growth

surface of the vessel. In this process, the

highly interactive gas plasma mixture is

exposed to ambient air. The consistency

of the treatment surface can therefore be

compromised by day-to-day environmental

changes.

At BD Biosciences, molded polystyrene

vessels are placed in a closed, highly-

controlled environment that ensures a

consistent treatment surface.2,3

BD Biosciences made a major research

investment to develop this unique vacuum-

gas plasma process, which

results in a hydrophillic surface on the

plastic that supports cell attachment and

spreading.4-6

This process is used to produce both

traditional tissue culture (TC) surfaces on

BD Falcon™ dishes, plates, flasks, and roller

bottles, as well as surfaces on BD Primaria™

cultureware. Recent research published in

Stem Cells demonstrates the superiority of

the BD Biosciences process.3

Cell performance tests ensure consistent results

A sensitive clonogenic assay85 using MRC-5

cells, a diploid human fibroblast line, is used

to validate the manufacturing process for

each BD Falcon TC product. Routine testing

of standard TC products is performed by

testing growth to confluency at 72 hours

with MRC-5 cells. The surface chemistry

of each lot of BD Primaria products is

confirmed by electron spectroscopy for

chemical analysis (ESCA).

Our Commitment to Quality

All BD Falcon, BD BioCoat, and

BD™ brand product manufacturing

sites are ISO 9000 certified. This

certification verifies that our facilities

meet international quality system

standards. The quality system is

routinely audited by a notified body

to ensure a work environment that

consistently maintains the highest

standards. ISO compliance gives our

customers an added level of assurance

that BD Biosciences is committed

to superior quality and continuous

product improvement. Copies of our

certificates can be obtained by calling

your local BD office.

Advanced Culture Surfaces for Neuronal Cells

Pseudo-colored merged image of Alexa Fluor® 488 (cell bodies and neurites: green) and Hoechst (nuclei: blue) channels of PC12 cells differentiated with 100 ng/mL NGF. Images were acquired as a 4x4 montage using a 20x objective (0.75 NA).



BD Falcon Flasks

description qty./pack qty./case cat.no.

CELLineTM 1000 System 1 3 353137

BD Falcon 75 cm2 standard TC Flask, vented cap 5 60 353136

BD Falcon 150 cm2 standard TC Flask, plug-seal cap 5 40 355000



BD Falcon Dishes


BD Falcon 35 mm Easy-Grip Style 20 500 351008Bacteriological Petri Dish

BD Falcon 35 mm Easy-Grip Style Cell Culture Dish 20 500 353001

BD Falcon 60 mm Center-Well Organ Cell Culture Dish 20 500 353037

BD Falcon 100 mm Standard Style Cell Culture Dish 20 500 353003

BD Falcon 150 mm Integrid Dish 10 100 351013

BD Falcon Cell Culture Plates


BD Falcon 6-well Multiwell Plate, flat-bottom 6 36 353224



BD Primaria 24-well Multiwell Plate, flat-bottom 1 50 353847


BD Falcon 96-well Microplates, TC, round-bottom 1 50 353077

BD Falcon 96-well Microplates, TC, flat-bottom 1 50 353070

BD Falcon 96-well Microplates, Standard, flat-bottom 1 50 351172

BD Falcon 96-well Microplates, Standard, round-bottom 1 50 351177

BD Falcon 384-well Optilux™ Microplates 5 50 353962

BD Falcon 1536-well Microplates 15 60 353249

BD Falcon Flasks

BD Falcon™ flasks are used to passage or to scale-up cells for

production. Vacuum-gas plasma tissue culture treatment provides

consistent cell attachment, spreading, and growth. Choose BD

Primaria™ non-treated polystyrene for standard tissue cultures or

BD BioCoat™ growth surfaces to meet your individual cell culture

requirements.

Visit bdbiosciences .com/flasks for a complete listing of BD Falcon flasks.

BD Falcon Cell Culture Plates

BD Falcon cell culture plates are available in well-characterized

dimensions and surface chemistries. These properties allow the use

of divergent cell culture methodologies and can be used for fluores-

cence, luminescence, colorimetric, and radiometric assays. Available

in configurations from 6- to 1536-well formats, these plates provide

the flexibility to meet the needs of a wide range of cell-based assays

and other biochemical assay needs in both manual and automated

settings.

BD Falcon Dishes

BD Falcon dishes are available in a wide range of sizes and

configurations to ensure they meet the needs of more

specialized applications.

The BD Falcon™ cultureware family also includes cell scrapers for

removing attachment cells from cultureware, cell strainers for

enhancing cell isolations from tissues, and various storage and

collection containers. Visit bdbiosciences .com/cellcultureware for

more information on these products.

Visit bdbiosciences .com/cellcultureplates for a complete listing of BD Falcon cell culture plates.

Visit bdbiosciences .com/cellculturedishes for a complete listing of BD Falcon cell culture dishes. Visit bdbiosciences .com/specialtydishes for more information on our bacteriological, compartment, and other specialty dishes.

BD Falcon Cultureware



BD Falcon Tubes


BD Falcon 50 mL Conical Tubes 25/bag 500 352070

25/rack 500 352098

BD Falcon 15 mL Conical Tubes 125/bag 500 352096

50/rack 500 352097

BD Falcon 5 mL Round-Bottom Tubes 125/pack 1000 352052



25/pack 500 352057

BD Falcon Pipets


BD Falcon 1 mL Serological Pipets 25/bag 1000 357506

100/box 1000 357521





BD Falcon Pipets

BD Falcon™ pipets, first in quality and first in innovation, are

manufactured at our ISO 9001-certified facility in a state-of-the-art,

high-performance work environment. Dedicated product teams use

advanced technologies and techniques, combined with rigorous

quality control procedures, to ensure the integrity of every pipet.

Each pipet component is made of the highest quality polystyrene

resin. Process control and attention to detail guarantee the finest

quality product. A critical volumetrics test verifies the accuracy of

each pipet by measuring the delivered volume. The final result is

consistently high-quality pipets that provide precise and accurate

liquid handling.

BD Falcon Tubes

Your life science research demands the most stable and controlled

environment possible for the analysis of biological and chemical

samples. At BD Biosciences, we manufacture our BD Falcon™

conical and round-bottom tubes from advanced bioanalytical-grade

resins. Our polymer selection process includes extensive testing to

ensure that the polymer does not leach unwanted substances.

BD Falcon tubes and our unique medical-style packaging provide

unsurpassed convenience and consistency.

BD Falcon Pipets and Tubes

Visit bdbiosciences .com/pipets for a complete listing of BD Falcon pipets.Visit bdbiosciences .com/tubes for a complete listing of BD Falcon tubes.

Cell Culture (continued)



Unique surface chemistry for enhanced cell culture

BD Biosciences has made a significant

research investment to develop a unique

vacuum-gas plasma process used to

produce both BD Primaria and traditional

tissue culture (TC) surfaces on BD Falcon

dishes, plates, flasks, and roller bottles.

The incorporation of nitrogen-containing

cations has been correlated to the attach-

ment and spreading of primary endothelial

cells in a clonal cell-growth assay.2 The

gases used to manufacture BD Primaria

cultureware contain both oxygen and

ammonia, resulting in the incorporation of

a variety of nitrogen-containing functional

groups on the surface, in addition to the

negatively charged oxygen-containing

groups found on traditional TC surfaces.

Because the complex surface on

BD Primaria cultureware is homogeneous

and stable, it has been used by researchers

for over a decade to improve attachment

and differentiation of a variety of cell types.

For example, cell biologists have used

BD Primaria cultureware for cultivating

neuronal cells,7 hepatocytes,8,9 and endo-

thelial cells.10 The surface chemistry of BD

Primaria products is confirmed by electron

spectroscopy for chemical analysis (ESCA).

BD Primaria Cultureware

When chick embryo spinal cord neurons are cultured on BD Primaria cultureware (left), growth is enhanced and extensive neurite development occurs. In this experiment, cells clumped and detached from traditional TC plates (above) after 20 days in culture, but remained viable and differentiated on BD Primaria cultureware.

Note: At pH 7, carboxy groups may be slightly dissociated and assume a negative (anionic) charge. Amine groups may protonate and assume a positive charge (cationic).

Traditional Tissue Culture Surface Chemistry BD Primaria™ Surface Chemistry

35 x 10 mm Easy-Grip Dish


Actual Dimensions: 40.28 mm O.D. x 6.17 mm

Actual Growth Area: 11.78 cm2

Working Volume: 2.5-3.0 mL 20 200 353801

60 x 15 mm Standard Dish





100 x 20 mm Standard Dish





25 cm2 Flask with Canted Neck


Nominal Growth Area: 25 cm2

Total Volume: 50 mL

Plug-Seal Caps 20 200 353813

Vented, 0.2 µm Caps 20 100 353808

75 cm2 Flask with Straight Neck


Nominal Growth Area: 75 cm2

Total Volume: 250 mL

Plug-Seal Caps 5 100 353824

Vented, 0.2 µm Caps 5 100 353810

6-well Multiwell Plate with Lid


Growth Area: 9.6 cm2

Working Volume: 15.5 mL 1 50 353846

24-well Multiwell Plate with Lid




96-well Multiwell Plate Flat-bottom with Lid






BD Primaria™ Cultureware (continued)




Extracellular Matrix Proteins and Attachment Factors

description qty. cat.no.

BD™ Extracellular Matrix, human 1mg 354237

BD Cell-Tak™ Cell and Tissue Adhesive 1mg 354240

5mg 354241

(2x5mg)10mg 354242

BD™ Osteopontin 50µg 354256

BD Collagen I, bovine 30mg 354231

BD Collagen I, human 250µg 354243

BD FIBROGEN Collagen I, human recombinant 250µg 354254

BD Collagen I, rat tail 100mg 354236

(10x100mg)1g 356236

BD Collagen I High Concentration (HC), rat tail 100mg 354249

BD Collagen II, bovine 5mg 354257

BD Collagen III, human 250µg 354244

BD FIBROGEN Collagen III, human recombinant 250µg 354255

BD Collagen IV, human 250µg 354245

BD Collagen IV, mouse 1mg 354233

(10x100µg)1mg 356233

BD Collagen V, human 250µg 354246

BD Collagen VI, human 500µg 354261

BD Fibronectin, human 1mg 354008

5mg 356008

(5x500mg)25mg 356009

BD Laminin, mouse 1mg 354232

BD Laminin/Entactin Complex High Concentration, mouse 10.5mg 354259

BD Ultrapure Laminin, mouse (entactin-free) 1mg 354239

BD Poly-D-Lysine 2mg 354210

BD Vitronectin, human 250µg 354238

FIBROGEN Human Recombinant Collagens are manufactured for BD Biosciences by FibroGen, Inc.

Visit bdbiosciences .com/ecms for more product information and technical references.

The development and normal functioning of cells depends on inter-actions with molecules in their microenvironment. The major classes of molecules that regulate cellular development and function include growth and differentiation factors, cell adhesion molecules, and the components of the extracellular matrix (ECM).

The ECM is a vital component of the cellular microenvironment in vivo. It serves as the framework for cell anchorage and plays a dynamic role in the regulation of cell growth and function.

The ECM exerts influences on behavior (adherence, spreading, differentiation, and migration) and the pattern of gene expression of the cells in contact with it. The ECM is not static but changes during both normal development and in tissue repair and regeneration; thus, is intimately involved in both normal biological function and response to injury.4

To create physiologically relevant in vitro models that support normal cell growth and function, the components of the in vivo environ-ment must be incorporated. Use of ECM as a coating for TC surfaces permits the development of model systems that closely mimic in vivo conditions.

Although the ECM of different tissues is specialized, the components commonly include collagens, fibronectin, laminins, proteoglycans, and vitronectin. Purified ECM proteins can be used in in vitro culture

systems to provide an appropriate environment for specific cell types and applications. The presence of ECM components promotes cell attachment, spreading, growth, and differentiation of a variety of primary cells and cell lines including neuronal stem cells. For some cells, the presence of particular ECM components is critical for optimum cell growth in vitro. Purified ECM proteins can be easily used to coat the desired tissue culture surface. Alternatively, BD BioCoat™ cellware, including tissue culture plates, dishes, flasks, coverslips, and cultureslides, are available pre-coated with a variety of ECM proteins and attachment factors. See additional BD BioCoat Cellware on pages 18–28.

Choice of ECM Substrata In Vitro

Nerve cells are more selective in their choice of substrata than most other cell types. Selection of an optimal substratum for neuronal cells in culture is often an empirical process. Primary neuronal cells exhibit minimal to no growth or differentiation on untreated glass or plastic surfaces. The choice of ECM or surface modification is an important component to consider when optimizing neuronal cell cultures. Recognizing the increasingly important role the ECM plays in the regulation of fundamental cellular processes, BD Biosciences offers a wide range of ECM proteins and attachment factors for researchers to incorporate into their cell culture systems.

Extracellular Matrix Proteins and Attachment Factors

*NA - not applicable †ND - not determined



• Promotes differentiation of many cell types

• Gels to form a three-dimensional model of basement membrane

• Increases rate of human tumor cell growth in nude mice

• Creates models to measure invasive potential of tumor cells

BD Matrigel™ basement membrane matrix

is a solubilized basement membrane

preparation extracted from the Engelbreth-

Holm-Swarm (EHS) mouse sarcoma, a tumor

rich in ECM proteins. Its major component

is laminin, followed by collagen IV, entactin,

and heparan sulfate proteoglycan.87-90 It also

contains TGF-b, fibroblast growth factor,

tissue plasminogen activator,91 and other

growth factors that occur naturally in the

EHS tumor.

At room temperature, BD Matrigel Matrix

polymerizes to produce a biologically active

matrix material resembling the mammalian

cellular basement membrane.

BD Matrigel Matrix is effective for the

attachment and differentiation of both

normal and transformed anchorage-

dependent cell types including neurons,

epithelial, and oligodendrocytes. These

include neurons, hepatocytes, Sertoli cells,

mammary epithelial cells, melanoma cells,

vascular endothelial cells, thyroid cells,

and hair folicle cells.88,92,94,95 BD Matrigel

basement membrane matrix will influence

casein gene expression in mouse mammary

epithelial cells.93,96 It also supports in vivo

peripheral nerve regeneration89 and

facilitates differentiation of bovine oviduct

epithelial cells.97

Growth Factor Reduced (GFR) BD Matrigel

Matrix is useful as an alternative to BD

Matrigel Matrix in many of the applications

listed above when a more highly defined

basement preparation is desired. It has

been used to define the signals necessary

in the formation of canicular cell processes

in bone cells,98 in elucidating the role of

growth factors in the formation of tubules

by primary mouse kidney cells,86 and in

gene expression studies of primary mouse

mammary epithelial cells.99 This phenol

red-free product is suitable for those assays

that require color detection (eg, colorimetric

assays and fluorescence detection).

All BD Matrigel basement membrane

matrices, including the High Concentration

formulation, can also be used to assess

in vivo angiogenic activity of different

compounds by subcutaneous injection

into mice (BD Matrigel Plug Assay).100-104

A similar approach can be used for the

transplantation and investigation of human

tumor cells. Reports include studies with

prostatic, breast, small-cell lung, colon,

adrenal carcinomas, melanomas, and

lymphoblastic leukemia cells.105-110

BD Matrigel Basement Membrane Matrix

Human Submandibular Gland (HSG) cells cultured on GFR BD Matrigel™ Matrix differentiate to form acinar structures within 24 hours. The acini shown were stained with H/E at 72 hours. Indirect immunofluorescence staining also revealed the salivary gland specific cysteine protease inhibitor, cystatin, in HSG cell acini (data not shown). (Photo courtesy of Dr. Hynda Kleinman.)

Amounts of Growth Factors (GF) Present in BD Matrigel™ Matrix vs. GFR BD Matrigel™ Matrix

Growth factor

Range of GF Concentration in

BD Matrigel MatrixAverage GF Concentration

in BD Matrigel MatrixTypical GF Concentration

in GFR BD Matrigel Matrix

EGF 0.5–1.3ng/mL 0.7ng/mL <0.5ng/mL

bFGF <0.1–0.2pg/mL NA* ND†

NGF <0.2ng/mL NA* <0.2ng/mL

PDGF 5–48pg/mL 12pg/mL <5pg/mL

IGF-1 11–24ng/mL 16ng/mL 5ng/mL

TGF-b 1.7–4.7ng/mL 2.3ng/mL 1.7ng/mL


BD Matrigel™ Basement Membrane Matrix


Formulation: Dulbecco’s Modified Eagle’s Medium with 10 µg/mL gentamycin. Typical protein concentrations are between 9-12 mg/mL. BD Matrigel Matrix is compatible with all culture media.

5mL 356234

10mL 354234

(5x10mL)50mL 356235

BD Matrigel Matrix High Concentration (HC)


Formulation: Dulbecco´s Modified Eagle´s Medium with 10 µg/mL gentamycin. Typical protein concentrations are between 18-22 mg/mL. BD Matrigel Matrix HC is compatible with all culture media.

10mL 354248

BD Matrigel Matrix, Phenol Red-free


Formulation: Dulbecco’s Modified Eagle’s Medium (without phenol red) with 50 µg/mL gentamycin. Phenol Red-free BD Matrigel Matrix is compatible with all culture media.

Standard Concentration 10mL 356237

High Concentration 10mL 354262

Growth Factor Reduced (GFR) BD Matrigel Matrix


Formulation: Dulbecco’s Modified Eagle’s Medium with 50 µg/mL gentamycin. GFR BD Matrigel™ Matrix is compatible with all culture media.

Purification: Purified by the method of Taub, et al., reducing the level of heparan sul-fate proteoglycan and several growth factors (eg, EGF, bFGF, IGF-1, PDGF, and NGF, but not TGF-b).86



High Concentration 10mL 354263

GFR BD Matrigel Matrix, Phenol Red-free


Formulation: Dulbecco’s Modified Eagle’s Medium (without phenol red) with 50 µg/mL gentamycin. Phenol Red-free BD Matrigel Matrix is compatible with all culture media.

Purification: Purified by the method of Taub, et al., reducing the level of heparan sul-fate proteoglycan and several growth factors (eg, EGF, bFGF, IGF-1, PDGF, and NGF, but not TGF-b). 86

10mL 356231

Dispase

Dispase is a bacillus-derived neutral metalloprotease that is recom-

mended for recovering cells cultured on BD Matrigel basement

membrane matrix. BD™ Dispase will yield a single cell suspension

far more gently and effectively than trypsin, collagenase, or other

proteolytic enzymes; it will not harm cells harvested for subcultiva-

tion or bioassays. In addition, BD Dispase may be used for tissue

dissociation. BD Dispase cleaves fibronectin, collagen IV, and to a

lesser extent collagen I, but it does not cleave collagen V or laminin.

Cell Recovery Solution

BD™ Cell Recovery Solution allows for the recovery of cells cultured

on BD Matrigel Basement Membrane Matrix for subsequent

biochemical analyses. BD Cell Recovery Solution depolymerizes

BD Matrigel Matrix gels without enzymatic digests and lengthy

incubation periods at high temperatures. Cells are released without

damage, thereby avoiding biochemical changes during incubation

and digestion of extracellular portions of cell-surface receptors and

adhesion molecules.

Cell Recovery Solution


BD Cell Recovery Solution 100mL 354253

BD Dispase


BD Dispase (5,000) caseinolytic units 100mL 354235

ECM Composition of BD Matrigel Matrix vs GFR BD Matrigel Matrix

Basement MembraneComponent

Percent in BD Matrigel Matrix

Percent inGFR BD Matrigel Matrix

Laminin 56% 61%

CollagenIV 31% 30%

Enactin 8% 7%





Pre-coated for Reproducibility and Convenience

BD BioCoat cellware combines the quality

of BD Falcon TC plates with a variety of

ECM proteins and attachment factors. The

result promotes cell attachment, spreading,

growth, and differentiation of a variety of

primary cells and cell lines in serum-free or

serum-containing cultures. A proprietary

manufacturing process applies the ECM

components to the vessel surface. The result

is a uniform, optically clear matrix substrate.

This technology, together with our exacting

quality control, guarantees the performance

of each lot, as well as consistency

from lot to lot.

Applications include:• Cell growth

• Cell adhesion assays

• Receptor-ligand binding assays

• Routine drug screening assays

• Studies of tissue morphogenesis

• Studies of cell-matrix interactions

• Regulation of signal transduction and gene expression

BD BioCoat Advantages

Ready-to-Use Convenience

Spend more time performing your

experiments rather than preparing for

them. Pre-coated BD BioCoat cellware

saves time and labor costs while

increasing productivity.

Quality Assurance Testing

Each lot of BD BioCoat cellware is

thoroughly tested for bioactivity and guar-

anteed to perform as claimed so you can

use them with complete confidence.

Reliable Performance

BD BioCoat cellware improves cell attach-

ment and increases proliferation rates for a

variety of normal and transformed cells.

Lot-to-Lot Consistency

BD Biosciences prides itself on maintaining

highly controlled ISO 9001 production

environments and validated manufacturing

procedures that result in uniformity and

consistent performance.

Wide Selection

Available with a wide range of ECM

proteins and attachment factors,

BD BioCoat cellware helps optimize

conditions for attachment, growth, or

differentiation for your cell type.

Custom Coatings

BD BioCoat Custom Coating Service

offers an extensive selection of cell-based

coatings on a wide variety of BD Falcon

vessels to meet your specialized needs.

BD BioCoat Cellware




Flasks are available in various sizes and designs with

standard, plug-seal, and vented cap options.

Multiwell plates are manufactured using a crystal-

grade polystyrene and feature patented labyrinth lid,

condensation rings, and deep-well design to control

contamination while reducing evaporation and mini-

mizing edge effects.

Dishes are exceptionally flat for distortion-free

optics and feature stacking rings for easier stacking

and handling.

CultureSlides have an innovative sealing design that

minimizes leakage and a plastic chamber affixed to a

specially cleaned glass slide that can be removed with

an easy-to-use, disposable safety removal tool.

BD BioCoat coverslips are composed of No.1 German

glass and provide an optically clear surface that is

non-neurotoxic and exhibits low background fluores-

cence. The convenient package also acts as a storage

container and allows for easy coverslip manipulation.

BD BioCoat coverslip-bottom dishes are 35 mm

dishes with a coverslip bottom that are easy to use

and facilitate preparation of cells for microscopic

analysis. The coverslip floor is No.1 German glass.

This format is ideal for use in high resolution and

inverted microscopy, fluorescence imaging in live

cells, confocal microscopy, phase contrast microscopy,

and micromanipulations.

BD Matrigel basement membrane matrix

is a solubilized basement membrane

preparation extracted from the Engelbreth-

Holm-Swarm (EHS) mouse sarcoma, a tumor

rich in ECM proteins. Its major component

is laminin, followed by collagen IV, heparan

sulfate proteoglycans, entactin, and

nidogen. BD Matrigel Matrix is effective for

the attachment and differentiation of both

normal and transformed anchorage depen-

dent cell types including neurons, epithelial,

and oligodendrocytes.

BD BioCoat Matrigel Matrix Cellware applications include:

• Elicitation of tissue-specific cellular morphology and protein production in epithelial cells

• Differentiation of neuronal, endothelial, and muscle cells

• Development of three-dimensional matrix model systems

BD BioCoat Matrigel Matrix has been used to culture:

• Rat hepatocytes26

• Primary human hepatocytes27

• Mouse pituitary gland tissue28

• Rabbit colonocytes29

• Human urothelial cells30

• Osteopontin (OPN) deficient rat vascular smooth muscle cells31

Source:• EHS mouse tumor

Formulation:• Dulbecco’s Modified Eagle’s Medium

with 50 μg/mL gentamycin

• BD Matrigel Matrix is compatible with all culture media

Quality Control:• Tested for ability to promote neurite

outgrowth from chick dorsal root ganglia in the absence of NGF

• Tested and found negative for bacteria and fungi

Storage and Stability:• Cellware stable for at least three

months at -20°C. Keep frozen until use.

• Thin layer cellware is stable for at least three months from date of shipment when stored at 2°C to 8°C.

Visit bdbiosciences .com/cellculturesystems

for more information on BD BioCoat matrigel

matrix cellware.

Multiwell Plates


6-well 2 354432

12-well 2 354503

24-well 2 354433

48-well 2 354508

Culture Dishes


35 mm 8 354460

Thin-Layer Multiwell and Assay Plates


6-well 5 354603

24-well 5 354605

96-well 5 354607

Thin-Layer Culture Dishes


35 mm 20 354602

60 mm 20 354601

100 mm 10 354600

For Hepatocytes


6-well plates 5 354510

100 mm culture dishes 5 354634

ES Culture Plates


6-well 5 354671

Rat Hepatocytes Cultured in the BD BioCoat Hepatocyte Differentiation Environment

Transmission electron micrograph of thin sections shows similar intracellular and intercellular

structures, indicative of healthy differentiated hepatocytes, in four-week-old cultures.

Intracellular structures active nucleus (5000x) Numerous mitochondria with calcium deposits; rough ER stacks near cell surfaces and surrounding mitochondria; glycogen stores; golgi, ribosomal rosette, lipid droplets.

Intercellular structures (8600x) Frequent interdigitation of apposing cells; gap and tight junctions; intercellular lumens with microvilli, characteristic of bile canaliculi.

BD BioCoat Matrigel Matrix Cellware






Laminin (LM), a major component of base-

ment membranes, is a multifunctional

glycoprotein that is used as a substrate to

culture and maintain differentiated func-

tion of a wide variety of cells. Laminin

has been shown in culture to stimulate

neurite outgrowth, promote cell attach-

ment, chemotaxis, cell differentiation, and

neuronal survival.

BD BioCoat Laminin cellware applications include:

• Promotion of cell attachment and spreading

• Induction of cell differentiation and neurite outgrowth

• Increase of proliferation of myoblasts32

• Study of effects of laminin on cell behavior


BD BioCoat Laminin cellware has been used to culture:

• SH-SY5Y (human neuroblastoma), Neuro-2A (mouse neuroblastoma), N1-E115 (rat neuroblastoma)33

• MCF-10A cells34,35

• SK-MEL-28 cells16

• HVSMC36

• MDA-231 breast cancer cell line21

Source:• EHS mouse tumor

Quality Control:• Tested for ability to initiate

differentiation (neurite outgrowth) of NG-108 rat glioma/mouse neuroblastoma cells


• Laminin purity >90% by SDS-PAGE (contains entactin)

Storage and Stability:• Stable for at least three months from

date of shipment when stored at 2°C to 8°C. Do not freeze.

Multiwell and Assay Plates


6-well 5 354404

12-well 5 354502

24-well 5 354412

48-well 5 354507

96-well 5 354410

Culture Dishes


35 mm 20 354458

60 mm 20 354405

100 mm 10 354452

150 mm 5 354553

Flasks


25 cm2 plug-seal† 10 354533

75 cm2 plug-seal†† 10 354522

† BD BioCoat 25 cm2 flasks are 70 mL canted neck

†† BD BioCoat 75 cm2 flasks are 250 mL canted neck

Effects of BD BioCoat Laminin Cellware on NG-108 Rat Glioma/Mouse Neuroblastoma Cells

NG-108 rat glioma/mouse neuroblastoma cells cultured on tissue culture plastic are loosely adhered and remain rounded.

NG-108 rat glioma/mouse neuroblastoma cells cultured on BD BioCoat Laminin cellware exhibit a spindle-shaped morphology and dendritic processes.

BD BioCoat Laminin Cellware

Type IV collagen is a ubiquitous component

in basement membranes and provides the

major structural support for this matrix.

When the collagen IV meshwork is assem-

bled, it provides a scaffold for the assembly

of other basement membrane components

through interactions with laminin, entactin/

nidogen, and heparan sulfate proteoglycan.

Collagen IV is useful as a substrate for

growth of nerve, epithelial, endothelial, and

muscle cells. Collagen plays a role in the

regulation of cell growth, differentiation

and adhesion, and tissue formation.

BD BioCoat™ Collagen IV cellware applications include:


• Cell differentiation and neurite outgrowth

• Increased proliferation of PC12 cells

• Studies of effects of collagen IV on cell behavior


BD BioCoat Collagen IV cellware has been used to culture:

• PC12 cells14,37,38

• SH-SY5Y cells14

• Human melanoma cells lines SK-MEL-28-N1 and SK-MEL-2816

• Primary murine hepatocytes39

Source:• EHS lathrytic mouse tumor

Quality Control:• Tested for ability to initiate

differentiation (neurite outgrowth) of NG-108 rat glioma/mouse neuroblastoma cells


• Collagen IV purity >90% by SDS-PAGE


date of shipment when stored at 2°C to 8°C.





6-well 5 354428

24-well 5 354430

96-well 5 354429

Culture Dishes


35 mm 20 354459

60 mm 20 354416

100 mm 10 354453

150 mm 5 354554

Flasks


25 cm2 plug-seal† 10 354534

75 cm2 plug-seal†† 10 354523

175 cm2 plug-seal 5 354528



Effects of BD BioCoat Collagen IV Cellware on PC12 Rat Pheochromocytoma Cells

PC12 cells cultured on tissue culture plastic do not attach well and tend to float in clumps in the culture medium.

PC12 cells cultured on BD BioCoat Collagen IV cellware show 90% attachment and rapid proliferation.

BD BioCoat Collagen IV Cellware




Poly-D-Lysine (PDL) and Poly-L-Lysine (PLL)

are synthetic compounds that enhance cell

adhesion and protein absorption by altering

surface charges on the culture substrate. In

addition to promoting cell adhesion, poly-

lysine surface treatments support neurite

outgrowth and improve the survival of

many central nervous system (CNS) primary

cells in culture. Because PDL and PLL are

synthetic molecules, they do not stimulate

biological activities that occur in vivo, and

they do not introduce impurities carried by

natural polymers.

BD BioCoat Poly-Lysine cellware applications include:

• Attachment and spreading of a variety of cell types

• Cell differentiation and neurite outgrowth

• Attachment of fastidious transfected cell lines

• Support survival of primary neurons in culture

• Serum-free or reduced serum culture

BD BioCoat Poly-Lysine cellware has been used to culture:

• Primary mouse brain capillaries40

• HEK-293 cells41-44

• MDA-231 breast cancer cells21

• Mouse cerebellar granule neurons45

• Transfected rat 1 cells46

• Rat anterior pituitary cells47

• Transfected COS-7 cells48

• Transiently transfected primary rat astrocytes49

• Rat primary cerebellar granule neurons50,51

• Murine microglia MG-7 cells52

Source:• PDL, synthetic (MW 75–150 kD)

• PLL, synthetic (MW 30–70 kD)

Storage and Stability:• Cellware stable for at least six months

from date of shipment when stored at 4°C to 30°C under dry conditions.

• Coverslips and culture slides stable for at least three months from date of shipment when stored at 2°C to 8°C.

Quality Control:• Multiwell plates (6, 12, 24, 48-well),

dishes, flasks, coverslips, and culture slides tested for ability to promote firm attachment of RCG cells.

• Assay plates (96- and 384-well) tested for signal from EcoPack™ 2-293 cells cultured in serum-free medium for one day, washed on a Skatron washer (Molecular Devices), and labeled with Calcein-AM. Intra- and inter-plate percent CVs are measured to ensure consistent coating. EcoPack 2-293 is a transformed HEK-293 cell line.

• All lots tested and found negative for bacteria and fungi.

Effect of BD BioCoat PDL on Cortical Neurons

Effect of BD BioCoat PDL on Rat Cerebellar Granule (RCG) Cells

Mixed culture of cortical neurons and astro-cytes cultured on BD BioCoat PDL cellware. Neurons are highly branched with very long processes. Astrocytes show similar process elongation.

RCG cells cultured on BD BioCoat PDL cell-ware show firm attachment (similar results obtained on PLL).

BD BioCoat Poly-Lysine Cellware

BD BioCoat Poly-D-Lysine Cellware Multiwell and Assay Plates

description qty. cat.no. qty. cat.no. qty. cat.no.

6-well 5 354413 50 356413

12-well 5 354470 50 356470

24-well 5 354414 50 356414

48-well 5 354509 50 356509

96-well Clear 5 354461 50 356461 80 356690

96-well Black/Clear 5 354640 50 356640 80 356692

96-well White/Clear 5 354651 50 356651 80 356693

96-well White 5 354620 50 356620 80 356691

384-well Black/Clear 5 354663 50 356663 80 356697

384-well White/Clear 5 354660 50 356660 80 356694

384-well Clear 5 354662 50 356662 80 356696

384-well White 5 354661 50 356661 80 356695

Coverslips (No . 1 German Glass)


12 mm Round 80 354086

Culture Dishes

description qty. cat.no. qty. cat.no.

35 mm 20 354467 100 356467

60 mm 20 354468 100 356468

100 mm 10 354469 40 356469

150 mm 5 354550

Coverslip-Bottom Dishes (No . 1 German Glass)


35 mm dish 20 354077with coverslip bottom

Flasks


25 cm2 plug-seal† 10 354479 50 356479

25 cm2 vented-cap† 10 354536 50 356536

75 cm2 plug-seal†† 5 354524 50 356524

75 cm2 vented-cap†† 5 354537 50 356537

150 cm2 plug-seal 5 354495 40 356495

150 cm2 vented-cap 5 354538 40 356538

175 cm2 plug-seal 5 354529 40 356529

175 cm2 vented-cap 5 354539 40 356539



CultureSlides


1-well 12 354566

2-well 12 354629

4-well 12 354577

8-well 12 354632

BD BioCoat Poly-L-Lysine Cellware Multiwell and Assay Plates


6-well 5 354515 50 356515

96-well Clear 5 354516 50 356516

Culture Dishes


35 mm 20 354518 100 356518

60 mm 20 354517 100 356517



12 mm Round 80 354085



BD BioCoat Poly-Lysine Cellware (continued)




For some applications, the use of a combi-

nation of ECM proteins, such as laminin

(LM) and human fibronectin (HFN), or LM

and attachment factors such as Poly-D-

Lysine (PDL) or Poly-L-Orinthine (PLO) has

been shown superior to the use of either

alone.

BD BioCoat™ PDL/LM and PLO/LM cellware

is suitable for culturing many different types

of peripheral nervous system (PNS) and

central nervous system (CNS) networks,

and is useful for promoting neural cell

attachment and differentiation. BD BioCoat

LM/HFN cellware provides an in vitro envi-

ronment that promotes cell attachment and

extensive process formation.

BD BioCoat PDL/LM, PLO/LM, and LM/HFN cellware applications include:

• Enhancement of neuronal cell attachment to plastic and glass

• Promotion of neurite outgrowth

• Culture of glial cells as a feeder layer for neurons

• Construction of neuronal cell model systems to study CNS function, development, and diseases

BD BioCoat PDL/LM has been used to culture:

• SH-SY5Y (human neuroblastoma), Neuro-2A (mouse neuroblastoma), N1-E115 (rat neuroblastoma)53

• Primary rat hippocampus54

• Murine T11-L3 dorsal root ganglion neurons (DRGNs)55,56

• Transfected PC12 cells56

• MCF-10A cells35

• Rat primary DRGNs57

Source:• PDL, synthetic (MW 75-150 kD)

• PLO, synthetic (MW 30-70 kD)

• LM, EHS mouse tumor

• HFN, human plasma

NOTE: Source material tested for hepatitis B antigen and HIV-1 antibody



Quality Control:• PDL/LM and PLO/LM tested for ability

to initiate differentiation (neurite outgrowth) of NG-108 rat glioma/mouse neuroblastoma cells


• LM/HFN tested for receptor agonist induced changes in intracellular calcium using Fluo-3A (a long wavelength fluorescent calcium indicator) in primary rat cortical neuron enriched cultures58,59

Multiwell and Assay PlatesPDL/LM


6-well 5 354595

24-well 5 354619

96-well 5 354596

Multiwell and Assay Plates (continued) PLO/LM


6-well 5 354658

24-well 5 354659

96-well 5 354657

LM/HFN


96-well 5 354670

Culture DishesPDL/LM


100 mm 10 354455

Coverslips (No . 1 German Glass)PDL/LM


12 mm round 80 354087

CultureSlidesPDL/LM


2-well 12 354687

8-well 12 354688

Effects of BD BioCoat LM/HFN on Primary Neurons

Neurotransmitter induced calcium released in rat primary neurons plated on BD BioCoat LM/HFN Cellware.

BD BioCoat Poly-D-Lysine/Laminin, Poly-L-Orinthine/Laminin, and Laminin/Human Fibronectin Cellware

Collagen I, found in most tissues and

organs, is most plentiful in dermis, tendon,

and bone. It is an integral part of the frame-

work that holds cells and tissues together

and has been recognized as a useful matrix

for improving cell culture. In vitro use of

collagen can exert effects on the adherence,

morphology, growth, migration, and differ-

entiation of a variety of cell types.11

BD BioCoat Collagen I cellware applications include:


• Rapid expansion of cell populations


• Cell adhesion assays improving survival of primary cells in culture

BD BioCoat Collagen I cellware has been used to culture:

• Primary murine cardiac myocytes12

• Human vascular SMC13

• PC12 cells and SH-SY5Y cells14

• Mouse primary keratinocytes15

• SK-MEL-28-N1 cells16

• Murine myoblast C2C12 cells17

• HUVEC18

• HEK-293 cells19

• Rat Kupffer cells20

• MDA-231 breast cancer cells21

Source:• Rat tail tendon

Quality Control:• Tested for ability to promote

attachment and spreading of HT-1080 human fibrosarcoma cells


• Collagen I purity >90% by SDS-PAGE

Storage and Stability:• Cellware stable for at least six months

from date of shipment when stored at 4°C to 30°C under dry conditions.

• Coverslips and CultureSlides stable for at least three months from date of shipment when stored at 2°C to 8°C.

CultureSlides


1-well 12 354556

2-well 12 354627

4-well 12 354557

8-well 12 354630



22 mm Round 60 354089

Flasks


25 cm2 plug-seal 10 354531 50 356531

25 cm2 vented-cap 10 354484 50 356484

75 cm2 plug-seal 5 354462 50 356462

75 cm2 vented-cap 5 354485 50 356485

150 cm2 plug-seal 5 354645 40 356645

150 cm2 vented-cap 5 354486 40 356486

175 cm2 plug-seal 5 354478 40 356478

175 cm2 vented-cap 5 354487 40 356487

Culture Dishes


35 mm 20 354456 100 356456

60 mm 20 354401 100 356401

100 mm 10 354450 40 356450

150 mm 5 354551


description qty. cat.no. qty. cat.no. qty. cat.no.

6-well 5 354400 50 356400

12-well 5 354500 50 356500

24-well 5 354408 50 356408

48-well 5 354505 50 356505

96-well Clear 5 354407 50 356407 80 356698

96-well Black/Clear 5 354649 50 356649 80 356700

96-well White/Clear 5 354650 50 356650 80 356701

96-well White 5 354519 50 356519 80 356699

384-well Black/Clear 5 354667 50 356667 80 356705

384-well White/Clear 5 354664 50 356664 80 356702

384-well Clear 5 354666 50 356666 80 356704

384-well White 5 354665 50 356665 80 356703

Please call for shipping on large orders

Effects of BD BioCoat Collagen I Cellware on Fetal Bovine Heart Endothelial (FBHE) Cells

FBHE cells grown for five days in basal medium containing 10% FBS on tissue culture plastic show sparse growth.

FBHE cells grown for five days using the BD BioCoat Endothelial Cell Growth Environment (Collagen I Cellware) form a confluent monolayer and show numerous mitotic cells.

BD BioCoat Collagen I Cellware




Human fibronectin (HFN) is a widely distrib-

uted glycoprotein that is used as a substrate

to promote attachment of cells through its

integrin-binding arginine-glycine-aspartate

(RGD) motif. HFN is a product of most

mesenchymal and epithelial cells and is

present in both the extracellular matrix

(ECM) and plasma. The principal function

of HFN appears to be in cellular migration

during wound healing and development,

regulation of cell growth and differentia-

tion, and haemostasis/thrombosis.

BD BioCoat Fibronectin cellware applications include:


• Rapid expansion of cell populations



• Study of effects of HFN on cell behavior

• Improve survival of primary cells in culture

BD BioCoat Fibronectin cellware has been used to culture:

• 3T3 Preadipocytes60

• Transfected 293T and transfected H1299 cells61

• MCF-10A cells34

• Primary cord blood mononuclear cells62

• SK-MEL-28 (human melanoma cells)16

• NIH3T3 cells63

• MDA-231 human breast cancer cells21

Source:• Human plasma

NOTE: Source material tested for hepatitis B antigen and HIV-1 antibody


attachment and spreading of BHK-21 hamster kidney cells


• Fibronectin purity >90% by SDS-PAGE





6-well 5 354402

12-well 5 354501

24-well 5 354411

48-well 5 354506

96-well 5 354409



Culture Dishes


35 mm 20 354457

60 mm 20 354403

100 mm 10 354451

150 mm 5 354552

Flasks


25 cm2 plug-seal† 10 354532

75 cm2 plug-seal†† 10 354521

150 cm2 plug-seal 5 354646

175 cm2 plug-seal 5 354526



22 mm round 60 354088

CultureSlides


1-well 12 354558

2-well 12 354628

4-well 12 354559

8-well 12 354631

Effects of BD BioCoat Fibronectin Cellware on BHK-21 Cells

BHK-21 fibroblasts cultured on glass CultureSlides do not spread.

BHK-21 fibroblasts cultured on BD BioCoat HFN CultureSlides attach and spread within one hour.

BD BioCoat Fibronectin Cellware





BD BioCoat™ Gelatin cellware provides

an attachment and growth-promoting

substrate for the culture of a variety of

cell types. Gelatin is used commonly in

the culture of vascular endothelial cells,

muscle, embryonic stem (ES) cells, and F9

teratocarcinoma cells. It is also suitable

for promoting adhesion of transfected cell

types. Gelatin is a heterogeneous mixture of

water-soluble proteins derived through the

hydrolysis of collagen.

BD BioCoat Gelatin cellware applications include:


• Culture of normal and transfected F9 teratocarcinoma cells for gene expression studies65

• Culture of human umbilical vein endothelial cells (HUVECs) for E-Selectin66 expression and VEGF induction

BD BioCoat Gelatin cellware has been used to culture:

• HUVEC,66 BME,22 BAEC23

• ES cells24

• C2C1235 and MM1464 myoblasts

• Normal and transfected F9 teratocarcinoma cells

Source:• Gelatin, porcine


proliferation of HUVECs


Storage and Stability:• Stable for at least six months from date

of shipment when stored at 4°C to 30°C under dry conditions.



6-well 5 354652 50 356652

96-well 5 354689 50 356689

Culture Dishes


100 mm 10 354653 40 356653

Culture Flasks


75 cm2 plug-seal† 5 354654 50 356654

75 cm2 vented cap† 5 354488 50 356488

†BD BioCoat 75 cm2 flasks are 250 mL canted neck

Effect of BD BioCoat Gelatin Cellware on Human Umbilical Vein Endothelial Cells (HUVECs)

HUVECs grown for seven days on BD BioCoat Gelatin 6-well multiwell plates seeded at a density of 2×104 in the presence of BD™ Endothelial Cell Culture Medium.

BD BioCoat Gelatin Cellware




BD BioCoat Custom Coating Service

Custom coating, the way you want it.

BD BioCoat Custom Coating Service offers

an extensive selection of cell-based coatings

on a wide variety of BD Falcon vessels—

from roller bottles to flasks to 384-well

plates. We can even develop special coat-

ings for you. Simply tell us the type of

vessel and coating you would like applied

and we’ll make a trial sample for you to

evaluate.

BD BioCoat Consulting Service

BD Biosciences is committed to helping you

optimize your cell culture conditions. The

process of selecting the most appropriate

substratum is often empirical and time

consuming. Our consulting service can

facilitate this process for you. Our highly

trained technical support staff will assist you

in determining which BD BioCoat product is

best for your cell type and application. Let

BD Biosciences put its comprehensive tech-

nical database and expertise in cell culture

to work for you.

BD BioCoat Bar-Coding Service

Get on the road to increased productivity

with barcoded BD BioCoat cellware. The

BD Bar-Coding Service provides high-quality

bar code labels affixed to any side of a

microplate. Bar codes have been quality

tested for optimal readability, chemical resis-

tance, and temperature durability.

BD Falcon and BD BioCoat cell culture inserts and insert systems

combine the microporous polyethylene terephthalate (PET)

membrane with multiple formats and pore sizes for a variety of

cell-based applications. A number of insert model systems have

been developed to study cells that are derived from the brain and

peripheral nervous system. For example, the BD Falcon FluoroBlok™

24-Multiwell Insert System has been used to examine dorsal root

ganglion neurite formation in an automated, fluorescence-based

assay.67 In addition to this application, BD cell culture insert products

are available for establishing co-culture systems and other models to

study the functional properties of neuronal and glial cells.

BD Falcon Cell Culture Inserts0.4 µm pore size


inserts for 6-well plates 1 48 353090



BD Falcon FluoroBlok Cell Culture Inserts


1.0 µm inserts for 24-well plates 1 48 351150



BD Falcon FluoroBlok 24-Multiwell Insert Systemswith Feeder Tray and Lid

description qty./pack cat.no.

1.0 µm pore size 1 351153

5 351154

BD Falcon FluoroBlok 24-Multiwell Insert Systemswith 24-well plate and lid



5 351156


5 351158

BD Falcon FluoroBlok 96-Multiwell Insert Systems



5 351162


5 351164

Visit bdbiosciences .com/cellcultureinserts for additional information about our cell culture inserts and insert systems.

BD BioCoatSM Services

Cell Culture Inserts and Insert Systems

Visit bdbiosciences .com/custom for additional information about our BD BioCoatSM Services.



Growth FactorsEpidermal Growth Factors


BD EGF, mouse natural (culture grade) 100µg 354001

(10x100µg) 100µg 356001

BD EGF, mouse natural (receptor grade) 100µg 354010

(5x100µg) 500µg 356010

BD EGF, human recombinant 100µg 354052

(10x100µg) 100µg 356052

Fibroblast Growth Factors


BD bFGF, bovine natural 10µg 356037

BD bFGF, human recombinant 10µg 354060

(5x10µg) 50µg 356060

(10x10µg) 100µg 356061

Insulin-like Growth Factor

BD IGF-I, human recombinant (culture) 10µg 354037

Nerve Growth Factors


BD 2.5S NGF, mouse natural 10µg 354005

100µg 356004

(2x500µg) 100µg 356005

BD 7S NGF, mouse natural 100µg 354009

Platelet-Derived Growth Factor


BD PDGF-BB, human recombinant 10µg 354051

(10x10µg) 100µg 356051

Vascular Endothelial Growth Factor


BD VEGF, human recombinant 10µg 354107

Transforming Growth Factor


BD TGF-b, human natural 1µg 354039

(5x1µg) 5µg 356039

(5x2µg) 10µg 356040

From purified growth factors to enriched culture supplements and

speciality media, these products all allow you to propagate your cells

under more defined serum-reduced or serum-free conditions.

BD Biosciences supplies a comprehensive line of high-quality cyto-

kines and media additives to meet your individual needs for culturing

animal or human cells in vitro:

• Vialed, highly purified growth factors with high biological activity

• Enriched culture supplements to be reconstituted in the medium of your choice

Lymphokines• Broad range of highly purified natural and

recombinant lymphokines

• Membrane filtered (0.2 μm membrane) and tested for biological activity

• High specific activity

• Available as vialed pure factors or as enriched conditioned media

Media Additives• Serum-free or low-serum alternatives to routinely used

high serum containing media

• Supplements and essential components used as the basis of specialized serum-free media

• Membrane filtered (0.2 μm membrane) and tested for biological activity

BD™ Growth Factors



LymphokinesInterleukin-1


BD IL-1b, human recombinant 2µg 354042

Interleukin-2


BD IL-2, human recombinant BRMP*units 10,000 354043

BRMP*units 50,000 356043

BD IL-2, rat natural BRMP*units 4,000 354110

BD IL-2, mouse recombinant BRMP*units 10,000 356078

BRMP*units 25,000 354078

Interleukin-3


BD IL-3, mouse recombinant 10µg 354058

Interleukin-4


BD IL-4, human recombinant 5µg 354068

Granulocyte-Macrophage Colony Stimulating Factor


BD GM/CSF, human recombinant 1µg 354048

Tumor Necrosis Factor-a


BD TNF-a, human recombinant 10µg 354066

(5x10µg) 50µg 356066

Stem Cell Factor


BD SCF, human recombinant 10µg 354105

Media Additives and SupplementsBD Nu-Serum Serum Replacements


BD Nu-Serum™ Serum Replacements 100mL 355100

500mL 355500

BD Nu-Serum™ IV Serum Replacements 100mL 355104

500mL 355504

BD Nu-Serum™ V Serum Replacements 100mL 355105

500mL 355505

Media Additives and Supplements (continued) T-Cell Culture Supplements


BD T-Cell Culture Supplement with ConA, rat (IL-2 culture supplement)

100mL 354115

BD T-Cell Culture Supplement without ConA, rat (IL-2 culture supplement)

100mL 354116

IL-3 Culture Supplement


BD IL-3 Culture Supplement, mouse 25mL 354040

Other Media Additives


BD Albumin, bovine serum (BSA, delipidized) 10g 354331

BD Linoleic Acid/Albumin Complex Linoleicacid/BSA 2.5/500mg 354227

BD Hydrocortisone 50mg 354203

BD Selenous Acid (sodium salt) 100mg 354201

BD Transferrin, human (holo) 100mg 354204

1g 354304

ITS Universal Culture Supplements


BD ITS Premix (5literequivalent) 5mL 354351

(20literequivalent) 20mL 354350

BD ITS+ Premix (2literequivalent) 20mL 354352

Mito+ Serum Extender


BD Mito+ Serum Extender (5literequivalent) 5mL 355006

Bovine Pituitary Extract


BD BPE 1.5mg 354123

(5x1.5mg) 7.5mg 356123


Endothelial Cell Growth Supplement

BD ECGS 1.5mg 354006

100mg 356006



* benzamide ribose monophosphate



Functional Assays and Discovery Reagents

Programmed cell death, or apoptosis, is a normal physiological

process that occurs during embryonic development and in the

maintenance of tissue homeostasis. The term apoptosis, from the

Greek word for the “falling off” of leaves from a tree, is used to

describe a process in which a cell actively participates in its own

destructive processes. The apoptotic program is characterized

by certain morphological features. These include changes in the

plasma membrane such as loss of membrane symmetry and loss

of membrane attachment, a condensation of the cytoplasm and

nucleus, and internucleosomal cleavage of DNA. In the final stages,

the dying cells become fragmented into “apoptotic bodies,”

which are rapidly eliminated by phagocytic cells without eliciting

significant inflammatory damage to surrounding cells. Inappropriate

induction of apoptosis has widespread pathological implications

and has been associated with diseases such as neurodegenerative

diseases, graft-versus-host diseases, transplant rejection, cancer,

and AIDS.73-77 The study of apoptosis has never been more important

than it is today. Along with this intensive study has come the

development of a multitude of methods to detect apoptotic cells

and dissect the mechanisms of apoptosis. It is important to note that

not every cell type will display all the classical features of apoptosis.

As such, studies designed to measure multiple aspects of apoptosis

may be required to analyze and confirm the mechanism of cell death.

BD Biosciences offers a full line of apoptosis detection research tools

and technologies for measuring indicators of apoptosis at different

stages. These reagents and technologies are based on the following

distinctive features of apoptosis:

• Plasma membrane alterations

• Mitochondrial changes

• Activation of caspases

• DNA fragmentation

For a complete list of over 400 apoptosis-related research reagents

and kits, visit bdbiosciences .com/literature to request our “Life,

Death, and Cell Proliferation” brochure.

Apoptosis



Cell Signaling*

description react clone isotype apps format size cat.no.

14-3-3 Hu C23-1 MouseIgG1 WB Purified 100µg 556596

14-3-3epsilon Chick,Dog,Frog,Hu,Ms,Rat 12 MouseIgM IP,IF,IHC,WB Purified 50µg 610542

IP,IF,IHC,WB Purified 150µg 610543

Akt (PKBa) Dog,Hu,Ms,Rat 2 MouseIgG2a IF,WB Purified 150µg 610877

7 MouseIgG1 IF,WB Purified 150µg 610837


Calreticulin Hu,Ms,Rat 16 MouseIgG1 IF,WB Purified 150µg 612137

CAPS Ms,Rat 4 MouseIgG1 WB Purified 50µg 612138

Caveolin (pY14) Hu,Ms,Rat 56 MouseIgG1 IF,IHC,WB Purified 150µg 611339

Caveolin 1 Chick,Hu 2234 MouseIgG2a IF,IP,WB Purified 150µg 610494

Chick,Dog,Hu,Ms,Rat 2297 MouseIgG1 FCM,IF,IHC,IP,WB Purified 150µg 610407

Dog,Hu,Ms,Rab,Rat C060 MouseIgM FCM,IF,IHC,IP,WB Purified 150µg 610058

Chick C20B MouseIgG1 IF,IP,WB Purified 150µg 610388

Caveolin 3 Ms,Rab,Rat 26 MouseIgG1 IHC,WB Purified 150µg 610421

cGB-PDE Rat 54 MouseIgG1 IF,WB Purified 50µg 611498

IF,WB Purified 150µg 611499

Dlg Hu,Ms,Rat 12 MouseIgG1 IF,IP,WB Purified 50µg 610874

IF,IP,WB Purified 150µg 610875

Doublecortin Ms 30 MouseIgG1 IF,WB Purified 50µg 611706

Gat Chick,Hu,Ms,Rat 3 MouseIgG1 IF,IHC,WB Purified 150µg 610589

GAP-43/Neuromodulin Hu,Ms,Rat 31 MouseIgG1 WB Purified 50µg 612262

WB Purified 150µg 612263

JBP1 (Skb1Hs) Dog,Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 150µg 611539

JIP-1 Ms,Rat 50 MouseIgG1 WB Purified 150µg 611891

Jun Bov,Chick,Dog,Hu,Ms,Rat 3 MouseIgG2a IF,IHC,IP,WB Purified 150µg 610327

N-Copine Ms,Rat 42 MouseIgG1 IF,WB Purified 50µg 611588

Dros 42 MouseIgG1 IF,WB Purified 50µg 612638

NCS-1 Hu,Ms,Rat 1 MouseIgG1 IF,WB Purified 50µg 611578

Basic classifications of protein kinases fall into these key areas of

research:

• G-protein coupled receptors

• G-protein regulators

• Intracellular hormone receptors

• Receptor tyrosine kinases

• Non-receptor tyrosine kinases

• Receptor serine/threonine kinases

• Non-receptor serine/threonine kinases

• Phospholipids and phospholipases

• Phosphatases

Signal transduction at the cellular level is the transference of external

cell signals to internal mechanisms. Simple signal transduction uses

receptor molecules such as the acetylcholine class: receptors that

constitute channels which allow signals to be passed in the form of

small ion movement, either into or out of the cell (see Ion Channels

and Transporter on page 50). Complex signal transduction relies on

the coupling of ligand-receptor interactions. Resulting events include

phosphorylations by tyrosine kinases and/or serine/threonine kinases.

Protein phosphorylations change enzyme activities and protein

conformations. The resulting alteration in cellular activity changes

the programming of gene expression and the up- and down-regu-

lation of mechanistic proteins. One of the key receptor classes in

neurobiology is tyrosine kinases (RTKs)—neurotrophin receptor

family (trkA, trkB, trkC) and NGF receptor.

Cell Signaling, Including Kinases and Phosphotases

*Visit bdbiosciences .com for a complete listing of cell signaling reagents.

Cell Signaling* (continued)


nNOS/NOS Type I Dog,Hu,Ms,Rat 16 MouseIgG2a IF,IHC,IP,WB Purified 50µg 610308

IF,IHC,IP,WB Purified 150µg 610309

Rat 52 MouseIgG1 WB Purified 50µg 611852


Hu,Ms,Rat Polyclonal RabbitIg IF,IHC,IP,WB Purified 50µg 610310


p116 Rip Dog,Hu,Ms,Rat 43 MouseIgG1 IF,WB Purified 50µg 611098

p38a (SAPK2a) Dog,Hu,Ms,Rat 27 MouseIgG1 IF,WB Purified 150µg 612169

p62dok Hu 45 MouseIgG1 WB Purified 150µg 610753

p96 (Disabled-2) Hu,Ms,Rat 52 MouseIgG1 IF,WB Purified 150µg 610465

Phocein Rat 38 MouseIgG2b IF,WB Purified 50µg 612694


PIN Hu,Rat 4 MouseIgG1 IF,IP,WB Purified 50µg 610726


PITPalpha Dog,Hu,Ms,Rab,Rat 50 MouseIgG1 IF,WB Purified 50µg 611024

PITPNM Ms,Rat 8 MouseIgG1 WB Purified 50µg 612026


RACK1 Bov,Chick,Dog,Frog,Hu,Ms,Rat 20 MouseIgM IF,IHC,IP,WB Purified 150µg 610178

Ras Chick,Dog,Hu,Ms,Rat 18 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610002

Ras-GRF2 Hu,Ms,Rat 23 MouseIgG1 IF,WB Purified 150µg 610841

SH2-B Hu,Ms,Rat 46 MouseIgG1 IF,WB Purified 50µg 611526


ShcC Hu,Ms,Rat 23 MouseIgG1 IF,IP,WB Purified 50µg 610642


SHPS-1 Rat 17 MouseIgG1 WB Purified 150µg 611497

SPA-1 Hu,Ms,Rat 3 MouseIgG1 IF,WB Purified 150µg 611215

Stat1 (C-Terminus) Chick,Dog,Hu,Ms,Rat 42 MouseIgG2b IF,IP,WB Purified 150µg 610186

Stat1 (N-Terminus) Chick,Dog,Frog,Hu,Ms,Rat 1 MouseIgG1 WB HRP 150µg 610118


Polyclonal RabbitIg IF,IP,WB Purified 150µg 610120

Stat1 (pY701) Hu,Ms 14 MouseIgG1 IF,IP,WB Purified 150µg 612133

4a MouseIgG2a IF,IP,WB Purified 150µg 612233

Stat2 Chick,Hu 22 MouseIgG2a IP,WB Purified 150µg 610188

Stat5 (MGF) Dog,Hu,Ms,Rat 89 MouseIgG2b FCM,IF,IHC,WB Purified 150µg 610192

Stat5 (pY694) Hu Polyclonal RabbitIg IF,WB Purified 150µg 611819

47 MouseIgG1 WB Purified 150µg 611965

Stat5A Dog,Hu,Ms,Rat 51 MouseIgG1 IF,WB Purified 150µg 611835

Stat6 (pY641) Hu 18 MouseIgG2a WB Purified 150µg 611567

Polyclonal RabbitIg WB Purified 150µg 611821

Striatin Dog,Hu,Ms,Rat 6 MouseIgG2b IF,WB Purified 50µg 610838

YT521-B Rat 8 MouseIgG1 IF,WB Purified 50µg 612380

ZO-1 Hu 1 MouseIgG1 IF,WB Purified 50µg 610966

Circadian Regulator


Per2 Ms,Rat 52 MouseIgG1 IF,WB Purified 50µg 611138

*Visit bdbiosciences .com for a complete listing of cell signaling reagents.



Functional Assays and Discovery Reagents (continued)

Kinases and Phosphatases


Calcineurin Chick,Frog,Hu,Ms,Rat 29 MouseIgG2a IF,IHC,WB Purified 150µg 610260

CaM Kinase Kinase Dog,Hu,Rat 6 MouseIgG2a IF,IHC,IP,WB Purified 50µg 610544


CaM Kinase II Dros 45 MouseIgG1 IF,IHC,WB Purified 50µg 612624

Dog,Hu,Ms,Rat 45 MouseIgG1 IF,IHC,WB Purified 50µg 611292

IF,IHC,WB Purified 150µg 611293

CaM Kinase IV Hu,Ms,Rat 26 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610275


CaM Kinase VI Ms,Rat 5 MouseIgG1 IF,WB Purified 50µg 611736


CASK Dog,Frog,Hu,Ms,Rat 7 MouseIgG1 IF,IP,WB Purified 50µg 610782


c-Raf-1 Chick,Dog,Hu,Ms,Rat 53 MouseIgG1 IF,IP,WB Purified 150µg 610152

CRIK Hu,Ms,Rat 6 MouseIgG1 IF,WB Purified 50µg 611376


DARPP-32 Ms,Rat 15 MouseIgG1 IF,WB Purified 50µg 611520

DGKi Hu,Ms,Rat 25 MouseIgG1 WB Purified 50µg 612150

Dyrk Hu,Rat 106 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610674


ERK (pan) Chick,Dog,Frog,Hu,Ms,Rat 16 MouseIgG2a IF,IHC,IP,WB Purified 150µg 610124

ERK1 Chick,Dog,Frog,Hu,Ms,Rat MK12 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610031

ERK2 Chick,Dog,Frog,Hu,Ms,Rat 33 MouseIgG2b IF,IHC,IP,WB Purified 150µg 610104

GSK-3b Chick,Dog,Hu,Ms,Rat 7 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610201


Dros 7 MouseIgG1 IF,IP,WB Purified 50µg 612634

GSK-3b (pY216) Hu,Ms,Rat 13a MouseIgG1 WB Purified 50µg 612312


Inhibitor 2 Hu 52 MouseIgG1 IF,WB Purified 150µg 610699

LIMK1 Hu,Ms,Rat 42 MouseIgG1 IF,WB Purified 150µg 611749

MRCKalpha Rat 41 MouseIgG1 IF,WB Purified 50µg 611584


MYPT1 Hu,Ms,Rat 20 MouseIgG1 WB Purified 150µg 612165

Ntk Chick,Dog,Frog,Hu,Ms,Rat 32 MouseIgG1 IF,IHC,WB Purified 150µg 610343

p38d (SAPK4) Hu 20 MouseIgG1 IF,WB Purified 150µg 610953

PDK1 (PKB Kinase) Dog,Hu,Ms,Rat 5 MouseIgG1 IF,WB Purified 150µg 611071

PKAC Dog,Hu,Ms,Rat 5B MouseIgG2b IF,WB Purified 150µg 610981

PKARI Chick,Dog,Frog,Hu,Ms,Rat 18 MouseIgG2b IF FITC 150µg 610168


PKARIa Chick,Dog,Hu,Ms,Rat 20 MouseIgG1 WB Purified 150µg 610610

PKARIIa Dog,Hu,Ms,Rat 40 MouseIgG1 IF,WB Purified 150µg 612243

PKARIIb Chick,Dog,Hu,Ms,Rat 45 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610626

PKCa Chick,Dog,Frog,Hu,Ms,Rat 3 MouseIgG2b IF,IHC,IP,WB Purified 150µg 610108

PKCd Hu,Ms,Rat 14 MouseIgG2b IF,IHC,WB Purified 150µg 610398

PKCe Chick,Dog,Hu,Ms,Rat 21 MouseIgG2a IF,IHC,IP,WB Purified 150µg 610086

PKCl Chick,Dog,Hu,Ms,Rat 41 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610208

PKCq Hu 27 MouseIgG2a IF,IP,WB Purified 150µg 610090

PKCb Chick,Hu,Ms,Rat 36 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610127




Kinases and Phosphatases (continued)


PKCg Ms,Rat 20 MouseIgG1 IF,WB Purified 50µg 611158

PKCiota Chick,Dog,Hu,Ms,Rat 23 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610175


Dros 23 MouseIgG2b IF,IHC,IP,WB Purified 50µg 612645

PP2A Catalytic a Dog,Hu,Ms,Rat 46 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610556

PRK1 (PKN) Dog,Hu,Ms,Rat 49 MouseIgG1 IF,IP,WB Purified 150µg 610687

PYK2 (CAK b) Chick,Dog,Frog,Hu,Ms,Rat 11 MouseIgG1 IF,IHC,IP,WB Purified 150µg 610549

Synaptojanin 1 Ms,Rat 26 MouseIgG1 WB Purified 50µg 612248


TAO1 Hu,Ms,Rat 22 MouseIgG2a IF,WB Purified 150µg 611369

Yes Chick,Dog,Hu,Ms,Rat 1 MouseIgG1 IF,IHC,WB Purified 150µg 610376




Phospho-specific Antibodies


aSynuclein (PY125) Hu I57-628 MsIgG2b,k WB Purified 100µg 558246

b-Dystroglycan (pY892) Hu 27.1 MsIgG1 IF,IHC,WB Purified 50µg 612524


Actopaxin (pS8) Hu J160-366 MsIgG1,k WB Purified 100µg 558374

Akt (pS472/pS473) (PKBa) Hu,Ms,Rat 104A282 MsIgG1 IP,WB Purified 50µg 550747

Akt (pS473) Hu J177-204.20 MsIgG1,k WB Purified 100µg 558368

Akt (pT308) Hu,Ms J1-223.371 MsIgG1,k WB Purified 100µg 558316

c-Cbl (pY700) Hu 47/c-Cbl(Y700) MsIgG1 IF,IHC,WB Purified 50µg 612304


c-Cbl (pY774) Hu 29/c-Cbl(Y774) MsIgG1 WB Purified 100µg 558035

c-Jun (pS63) Hu 2 MsIgG1 IHC(F),WB Purified 100µg 558036

Caveolin (pY14) Hu,Ms,Rat 56 MsIgG1 IF,IHC,WB Purified 50µg 611338


Caveolin 2 (pY27) Hu 40/Caveolin2 MsIgG1,k WB Purified 100µg 558364

CD22 (BL-CAM) (pY828) Hu 46 MsIgG1 IHC,WB Purified 100µg 558029

CD22 (BL-CAM) (pY843) (BL-CAM) Hu 12a MsIgG1 IHC,WB Purified 100µg 558030

CD117 (pY568/pY570) (c-Kit) Hu K39-686 MsIgG1,k WB Purified 100µg 558390

CD221 (pY950) (IGF-1 Receptor) Hu J95-626 MsIgG2b,k WB Purified 100µg 558373

Cdk1/Cdc2 (pY15) Hu 44 MsIgG1 IHC,WB Purified 50µg 612306

IHC,WB Purified 150µg 612307

CREB (pS133) Hu J151-21 MsIgG1,k WB Purified 100µg 558359

CrkL (pY207) Hu K30-391.11.30 MsIgG2a,k WB Purified 100µg 558386

EGF Receptor (pY845) Hu 12A3 MsIgG1,k WB Purified 100µg 558381

EGF Receptor (pY1173) Hu 9H2 MsIgG1,k WB Purified 100µg 558382

EGF Receptor, Activated Form Hu 74 MsIgG1 IF,IP,WB Purified 50µg 610025


ERK1/2 (pT202/pY204) (p44/42 MAPK) Hu,Ms,Rat 20A MsIgG1 IHC,WB Purified 50µg 612358


A key area of neuroscience is the study of intracellular communi-

cation. Binding of ligands to receptors initiates a phosphorylation

signal cascade mediated via kinases and phosphatases (localized

in the cell membrane, cytoplasm, and nucleus). Phosphorylation of

tyrosine, serine, and threonine residues is critical for the control of

protein activity involved in cellular events. A myriad of kinases and

phosphatases regulates intracellular protein phosphorylation in many

different cell signaling pathways, such as those regulating apoptosis,

growth and cell cycle control, neurotransmitter regulation, and stress

responses.

Historically, phosphorylation detection has been performed using

techniques such as radiometric kinase assays and phosphoamine

acid labeling. In recent years, mass spectrometry techniques have

also been developed, but have a tendency to be time consuming

and cumbersome. However, the advent of phospho-specific anti-

bodies has facilitated the use of more straightforward techniques

such as WB, IP, and IF. BD Biosciences, in collaboration with Garry

Nolan, PhD at Standford University, has developed the

BD™ Phosflow technique. This technology combines phospho-

specific antibodies with the power of flow cytometry to enhance

phosphoprotein studies by reducing sample size and increasing

quantitative analysis, while allowing multiparameter analysis of

single cells and distinct cell subpopulations. This approach has been

extended using BD™ Cytometric Bead Arrays (CBA) to analyze

protein phosphorylation in cell and tissue lysates. BD Biosciences

offers a variety of phospho-specific antibodies for a variety of experi-

mental techniques, including western blotting, immunoparticipation,

immunohistochemistry, immunofluorescence microscopy, and flow

cytometry.

The reagents, instruments, and tools that BD Biosciences offers are

designed to facilitate investigation of the signal transduction and

apoptotic pathways linking cellular components and processes to

one another during development, homeostasis, and disease.

Spotlight on Phoshoprotein Analysis



Phospho-specific Antibodies (continued)


Ezrin (pT567) Hu J37-954.281.307 MsIgG1,k WB Purified 100µg 558357

Ezrin (pY353) Hu I66-386 MsIgG1 IHC,WB Purified 100µg 558033

FADD (pS194) Hu J119-857.36 MsIgG1,k WB Purified 100µg 558370

FAK (pY397) Hu,Ms 18 MsIgG1 IF,WB Purified 50µg 611806


FAK (pY397) Hu 14 MsIgG1 IF,IHC,WB Purified 50µg 611722


Fyn (pY528)/c-Src (pY530) Hu 31 MsIgG2b IHC,WB Purified 50µg 612668


GSK-3b (pY216) Hu,Ms,Rat 13a MsIgG1 IHC,WB Purified 50µg 612312


IkBa (pS32/pS36) Hu 39A1431 MsIgG1 WB Purified 50µg 551818

IRS-1 (pY896) Hu K9-211 MsIgG2a,k WB Purified 100µg 558378

JNK (pT183/pY185) Hu 41 MsIgG1 IHC,WB Purified 50µg 612540


MARCKS (pS152/pS156) Ms I84-1233 MsIgG1,k WB Purified 100µg 558380

MEK1 (pS298) Hu,Ms J114-64 MsIgG1,k WB Purified 100µg 558375

Neurofilament NF-H Rat RNF405 MsIgM WB Purified 50µg 551958

RNF404 MsIgG2a WB Purified 50µg 551348

Neurofilament NF-M Rat RNF406 MsIgG1 WB Purified 50µg 551962

NF-kB (pS529) Hu K10-895.12.50 MsIgG2b,k WB Purified 100µg 558393

NF-kB p65 (pS536) Hu J144-460 MsIgG1,k WB Purified 100µg 558377

p38 MAPK (pT180/pY182) Hu,Ms,Rat 36 MsIgG1 IF,IHC,WB Purified 50µg 612288


30 MsIgG1 IF,IHC,WB Purified 50µg 612280


p53 (pS37) Hu J159-641.15.4 MsIgG1,k WB Purified 100µg 558369

p53 (pS46) Hu I117-1091 MsIgG1,k IHC,WB Purified 100µg 558245

p90 RSK1 (pS380) Hu 20a MsIgG1 IHC,WB Purified 50µg 612692


p120 Catenin (pY228) Hu 21a MsIgG1 IF,IHC,WB Purified 50µg 612536


p130Cas (pY249) Hu J169-757.12.2 MsIgG2b,k WB Purified 100µg 558401

Paxillin (pY31) Hu 19/Paxillin(Y31) MsIgG1,k WB Purified 100µg 558356

Paxillin (pY118) Hu,Ms 30 MsIgG1 IHC,WB Purified 50µg 611724


PDPK1 (pS241) (PDK1) Hu J66-653.44.22 MsIgG1,k WB Purified 100µg 558395

PLK1 (pT210) Hu K50-483 MsIgG1,k WB Purified 100µg 558400

Progesterone Receptor (pS190) Hu 1154/F12 MsIgG1,k WB Purified 100µg 558387

Rb (pS780) Hu J146-35 MsIgG1,k IHC,WB Purified 100µg 558385

Rb (pS807/pS811) Hu J112-906 MsIgG1,k WB Purified 100µg 558389

Rb, Underphosphorylated (a.a. 514-610) Hu G99-549 MsIgG1 IHC,IP,WB Purified 100µg 554164

Stat1 (pY701) Hu,Ms 14 MsIgG1 IF,IP,WB Purified 50µg 612132


4a MsIgG2a IF,IHC,IP,WB Purified 50µg 612232


Stat2 (pY690) Hu 7a/Stat2(pY690) MsIgG1,k IHC,WB Purified 100µg 558095




Phospho-specific Antibodies (continued)


Stat3 (pS727) Hu 49 MsIgG1 IF,IHC,WB Purified 50µg 612542


Stat3 (pY705) Hu,Ms 4 MsIgG2a IHC,WB Purified 50µg 612356


Stat4 (pY693) Hu 38/p-Stat4 MsIgG2b IHC,WB Purified 50µg 612738


Stat5 (pY694) Hu Polyclonal RabIg IF,WB Purified 50µg 611818


47 MsIgG1 IHC,WB Purified 50µg 611964


Stat6 (pY641) Hu Polyclonal RabIg IF,WB Purified 50µg 611820


18 MsIgG2a IF,IHC,WB Purified 50µg 611566


Ms J71-773.58.11 MsIgG1,k WB Purified 100µg 558241

Tyk2 (pY1054/pY1055) Hu I114-617 MsIgG1,k WB Purified 100µg 558394



BD Phosflow Reagents*

description clone apps format size cat.no.

AKT (pS473) F29-763 IC/FCM AlexaFluor®647 50tests 558314

AKT (pT308) J1-223.371 IC/FCM PE 50tests 558275

BLNK (pY84) J117-1278 IC/FCM AlexaFluor®488 50tests 558444

IC/FCM AlexaFluor®647 50tests 558443

IC/FCM PE 50tests 558442

Btk (pY551) & Itk (pY511) 24a/BTK(Y551) IC/FCM PE 50tests 558129



c-Cbl (pY700) 47/c-Cbl(Y700) IC/FCM PE 50tests 558087



c-Cbl (pY774) 29/c-Cbl(Y774) IC/FCM PE 50tests 558102


Caveolin (pY14) 56 IC/FCM PE 50tests 612568

CD140b (pY771) J23-1044 IC/FCM PE 50tests 558426











CD247 (pY142) K25-407.69 IC/FCM PE 50tests 558448



CREB (pS133) J151-21 IC/FCM PE 50tests 558436



EGF Receptor (pY845) 12A3 IC/FCM AlexaFluor®647 50tests 558523

EGF Receptor (pY1173) 9H2 IC/FCM AlexaFluor®647 50tests 558524

BD Phosflow is a highly innovative flow cytometry based technology,

enabling activation state analysis of multiple proteins at single-cell levels.

Using phosphorylation site specific antibodies in combination with

cell surface markers, you can study phosphorylation events directly in

small subpopulations of complex primary samples.

This revolutionary technology will not only enhance cell signaling

network analysis and high content secondary screening for drug

discovery and development, but also will enable the identification of

signaling pathway dependent biosignatures for disease.

BD Phosflow Reagents BD Phosflow Advantages

• Collect data for individual cells

• Analyze rare cell types directly in complex primary samples

• Correlate multiple phosphorylation events simultaneously

• Perform rapid and scalable screens in 96-well plates using

BD FACS™ flow cytometers with the BD™ HTS option



*Visit bdbiosciences .com/phosflow for a complete listing of BD Phosflow reagents.


BD Phosflow Reagents (continued)


ERK1/2 (pT202/pY204) 20A IC/FCM PE 50tests 612566



ERK2 G263-7 IC/FCM AlexaFluor®488 50tests 558525


FAK (pS910) K73-480 IC/FCM PE 50tests 558540


Histone H3 (pS28) HTA28 IC/FCM AlexaFluor®647 50tests 558217

IRS-1 (pY896) K9-211 IC/FCM PE 50tests 558440


JNK (pT183/pY185) Polyclonal IC/FCM,IHC(F),WB Purified 50tests 558268

LAT (pY171) I58-1169 IC/FCM PE 50tests 558487



LAT (pY226) J96-1238.58.93 IC/FCM PE 50tests 558433



Lck (Y505) 4/Lck-Y505 IC/FCM PE 50tests 558552


MEK1/2 (pS222) Polyclonal IC/FCM,IHC(F), Purified 50tests 558281 IHC(F)†,WB

NF-kB p65 (pS529) K10-895.12.50 IC/FCM PE 50tests 558423



Notch1 mN1A FCM,IC/FCM,WB PE 100µg 552768

NPM-ALK ALK1 IC/FCM FITCSet 100tests 559818

IC/FCM PESet 100tests 559257

p38 MAPK (pT180/pY182) 36 IC/FCM PE 50tests 612565



p120 Catenin (pS288) 17/catenin IC/FCM AlexaFluor®647 50tests 558561

p130Cas (pY249) J169-757.12.2 IC/FCM AlexaFluor®488 50tests 558546


PLC-g1 (Y783) 27 IC/FCM AlexaFluor®488 50tests 557884


PLC-g2 (pY759) K86-689.37 IC/FCM PE 50tests 558490



PLK1 (pT210) K50-483 IC/FCM PE 50tests 558445



Rb (a.a. 332-344) G3-245 IC/FCM FITCSet 100tests 556538


Rb (pS780) J146-35 IC/FCM AlexaFluor®488 50tests 558554


Rb, underphosphorylated (a.a. 514-610) G99-549 IC/FCM FITCSet 100tests 550501


†Antigen retrieval required




BD Phosflow Reagents* (continued)


SLP-76 (pY128) J141-668.36.58 IC/FCM PE 50tests 558437



Stat1 (pY701) 4a IC/FCM PE 50tests 612564



Stat3 (pS727) 49/p-Stat3 IC/FCM AlexaFluor®488 50tests 558085


Stat3 (pY705) 4 IC/FCM PE 50tests 612569

FCM,WB AlexaFluor®488 50tests 557814

FCM,WB AlexaFluor®647 50tests 557815

Stat4 (pY693) 38/p-Stat4 IC/FCM PE 50tests 558249









J71-773.58.11 IC/FCM PE 50tests 558252



Syk (p352) Pan/Phospho 4D10 IC/FCM FITC 50tests 557952

Syk (pY348) I120-722 IC/FCM PE 20µL 558529

TCL1 H29-1349 IC/FCM PE 50tests 558025

Zap70 (pY292) J34-602 IC/FCM PE 50tests 558510



Zap70 (Y319)/Syk (Y352) 17a IC/FCM PE 50tests 557881



BD Phosflow Supporting Reagents

description clone isotype format size cat.no.

BD Cytofix Fixation Buffer 100mL 554655

BD Phosflow Fix Buffer 250mL 557870

BD Phosflow Lyse/Fix Buffer (5X) 250mL 558049

BD Phosflow Perm/Wash Buffer I 125mL 557885

BD Phosflow Perm Buffer II 125mL 558052

BD Phosflow Perm Buffer III 125mL 558050

Staining Buffer (FBS) 500mL 554656

Mouse IgG1 Isotype Control MOPC-21 MsIgG1 AlexaFluor®488 50tests 557782

AlexaFluor®647 50tests 557783

PE 50tests 551436

Mouse IgG2a Isotype Control MOPC-173 MsIgG2a PerCP-Cy™5.5 50tests 558020





Cytoskeletal Proteins


4.1N Chick,Dog,Hu,Ms,Rat 4 MouseIgG1 IF,WB Purified 50µg 611836

Actin Chick 4 MouseIgG1 IF,WB Purified 50µg 612656


Arp3 Chick,Dog,Hu,Ms,Rat 4 MouseIgG1 IF,WB Purified 50µg 612134

150µg 612135

BART Hu,Ms,Rat 6 MouseIgG1 WB Purified 50µg 612036


CLIP-115 Hu,Ms,Rat 14 MouseIgG1 IF,WB Purified 50µg 611142


Cofilin Dog,Hu,Ms,Rat 32 MouseIgG1 WB Purified 50µg 612144

150µg 612145

Collybistin Ms,Rat 3 MouseIgG1 IF,WB Purified 50µg 612076

Contactin Hu,Ms,Rab,Rat 41 MouseIgG1 IF,IP,WB Purified 50µg 610578


Dystrobrevin Ms,Rab,Rat 23 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610766

Rab,Rat 38 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610780


The cytoskeleton regulates cell shape,

transport, motility, and integrity.

Cytoskeleton consists of filament systems

with a large number of associated proteins

that regulate each system. Cytoskeletal

filaments that are found in all cells can be

grouped into microfilaments, intermediate

filaments, and microtubules.

Microfilaments Microfilaments are the smallest (7–9 nm in diameter) of the three cytoskeletal filaments. They consist of actin and tropomyosin in association with a large number of other proteins.

• Mammalian actin exists in six different forms (isoforms): four associated with different types of muscle and two cytoskeletal isoforms found in all non-muscle cells.

- cytoskeletal isoforms are b and g- muscle isoforms are a (skeletal), b

(cardiac), g (smooth), and g (enteric)

• Actin is usually the most abundant protein in cells.

• It is 375 amino acids in length and isoforms can be separated on two-dimensional protein gels.

• b and g cytoskeletal actin differ by four amino acids at their amino end.

• Actin can be identified in its monomer (g-actin, globular) or polymer (f-actin, filamentous) form.

• Actin can also be identified by antibodies to all, or specific, isoforms.

• Actin mRNA is localized to specific regions within the cell cytoplasm.

Intermediate FilamentsIntermediate filaments are rope-like fibers made of eight tetramers, 10 nm in diameter. They are intermediate in size between microfilaments and microtubules. Different cell types express different combinations of intermediate filaments.

• Their main function is to maintain cell integrity. Most cells express more than one intermediate filament protein.

• The nucleus has its ownintermediate filament network (lamins).

• Some examples are keratin (skin),neurofilament (neurons), and glial fibrillary acid protein (GFAP, glia).

MicrotubulesLargest of the three filaments, 24 nm in diameter, microtubules form hollow tubes made of 13 parallel proto-filaments, involved with cytoplasmic transport of organelles, proteins, and vesicles.

• Different motors move in different directions on microtubules.

• Tubules composed of heterodimers of tubulin. There are two isoforms of tubulin: b and g.

• Stabilized by a large family of different microtubule associated proteins (MAPs). The most famous MAP is tau, found in neurofibrillary tangles associated with Alzheimer’s Disease (AD).78

Cytoskeletal Proteins



Cytoskeletal Proteins (continued)


GFAP Bov,Chick,Dog,GPig, 1B4 MouseIgG2b IF,IHC(F),WB Purified 500µg 556328 Hu,Ms,Pig,Rab,Rat,Sheep

2E1 MouseIgG2b IF,IHC(F),WB Purified 500µg 556329

4A11 MouseIgG2b IF,IHC(F),WB Purified 500µg 556327

Chick,Hu,Ms,Rat 52 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610565


GFAP Cocktail Bov,Chick,Dog,GPig, 4A11,1B4,2E1 MouseIgG2b IF,IHC(F),WB Purified 500µg 556330 Hu,Ms,Pig,Rab,Rat,Sheep

KAP3A Chick,Dog,Hu,Ms,Rat 14 MouseIgG1 IF,IP,WB Purified 50µg 610637

Katanin p80 Hu 19 MouseIgG1 IF,WB Purified 150µg 611411

KIF1A Ms 16 MouseIgG1 IF,WB Purified 50µg 612094

KIF3A Hu,Ms,Rat 28 MouseIgG1 IF,WB Purified 150µg 611509

Laminin B2 Hu,Rat 22 MouseIgG1 IF,IP,WB Purified 50µg 610722

MAP1B Ms 6 MouseIgG2a WB Purified 50µg 612678


MAP1B (LC1) Ms 19 MouseIgG1 WB Purified 50µg 612680


MAP2 Bov,Frog,Hu,Qua,Rat Ap20 MouseIgG1 IHC(Fr),IP,WB Purified 100µg 556320

MAP2B Hu,Ms,Rat 18 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610460

MKLP1 Hu 24 MouseIgG2a WB Purified 50µg 612682


Nestin Hu,Ms,Rat 25 MouseIgG1,k IF,WB Purified 50µg 611658


Rat Rat401 MouseIgG1 EM,FCM,IF, Purified 100µg 556309 IHC(P),WB

Neurofilament NF-H Rat RNF402 MouseIgM WB Purified 50µg 551349

RNF404 MouseIgG2a WB Purified 50µg 551348

RNF405 MouseIgM WB Purified 50µg 551958

Neurofilament NF-M Rat RNF403 MouseIgG1 WB Purified 50µg 551957

RNF406 MouseIgG1 WB Purified 50µg 551962

Ninjurin Dog,Hu,Ms,Rat 50 MouseIgG2a IF,IHC,WB Purified 50µg 610776

150µg 610777

Occludin Dog,Hu,Ms,Rat 19 MouseIgG1 WB Purified 50µg 611090

150µg 611091

p150 Glued Chick,Dog,Hu,Ms,Rat 1 MouseIgG1 IF,IP,WB Purified 50µg 610473


Rat 12 MouseIgG1 IF,WB Purified 50µg 612708


p24 Ms,Rat 22 MouseIgG1 IF,WB Purified 50µg 611616


Tau Rat 15 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610672


TAU-5 MouseIgG1 IHC(F),WB Purified 100µg 556319

b-Tubulin Bov,Hu,Ms,Rat 5H1 MouseIgM ICC,WB Purified 100µg 556321

VAP33 Dog,Hu,Ms,Rat 8 MouseIgM IF,WB Purified 50µg 612180






Development and Differentiation


d-Catenin Ms,Rat 30 MouseIgG1 WB Purified 50µg 611536

150µg 611537

ADNP Hu 45 MouseIgG1 IF,WB Purified 50µg 612512

BERP Ms,Rat 27 MouseIgG1 IF,WB Purified 50µg 611732


beta -Dystroglycan Hu 56 MouseIgG2a IF,WB Purified 50µg 612090


beta -Dystroglycan (pY892) Hu 27.1 MouseIgG1 FCM,IF,WB Purified 50µg 612524

FCM,IF,WB Purified 150µg 612525

Brevican Dog,Ms,Rat 2 MouseIgG1 IF,WB Purified 50µg 610894


CD171 Hu 5G3 MouseIgG2a IP Purified 100µg 554273

CD56 Ms 12F8 RatIgM IHC(Fr),WB Purified 100µg 556325

12F11 RatIgG2a IHC(F),IHC(Fr), Purified 100µg 556323 WB,IHC(P)

Ms,Rat N-CAM13 MouseIgG2a IHC(Fr),WB Purified 100µg 556324

CRMP5 Ms 8 MouseIgG1 IF,WB Purified 50µg 612364

Doublecortin Ms 30 MouseIgG1 IF,WB Purified 50µg 611706

DP103/Gemin3 Hu 2 MouseIgG2b WB Purified 50µg 612152

150µg 612153

ENC-1 Dros 7 MouseIgG1 IF,IHC,WB Purified 50µg 612628

Hu,Ms,Rat 7 MouseIgG1 IF,WB Purified 50µg 611210

FAK Chick,Dog,Hu,Ms,Rat 77/FAK MouseIgG1 IF,IHC,IP,WB Purified 50µg 610087

150µg 610088

FAK (pY397) Hu,Ms 14 MouseIgG1 IF,WB Purified 150µg 611723


Fyn Dog,Hu,Rat 25 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610163


Fyn (pY528)/c-Src (pY530) Hu 31 MouseIgG2b WB Purified 50µg 612668


Galpha o Hu 23 MouseIgG2a IF,WB Purified 50µg 612526


For most species, the nervous system is the most complex part of

the organism. For instance, in mammalian species, over one third of

the genome is expressed in the nervous system. The development

process is complex and requires the precise control of many events

that generate a large variety of cell types with absolute correct

positioning and interconnections. The development of the nervous

system starts early in embryonic life and creates and maintains the

complex system of structures—which includes the brain, spinal

cord, and peripheral nerves—throughout the life of the individual.

Very early in the embryonic life, the neural plate and neural tube

are induced and the neural precursors form and proliferate. These

precursors will develop into neurons or glial cells. Neuronal and

glial precursors migrate from their site of origin to final positions in

the region they will occupy. They will then differentiate into mature

phenotypes, expressing appropriate functional molecules, extending

axons, setting up synaptic connections, and undergoing myelination

and additional modifications to delineate mature structures. During

the process, many neurons undergo programmed cell death.

Some key processes and molecules that guide development are:

• The effects of morphogens and Hox genes in pattern formation and the specification of cell fate

• Cell-to-cell interactions that regulate specification, and the molecules that effect those, such as notch and delta interactions

• Boundary regions that contain specialized cells such as “organizers”

• Growth in cortex regulated and guided by glial cells

• The effects of neurotransmitters

• The role of genetic programs in the formation of synapses versus activity-regulated remodeling of neuronal connections

Development and Differentiation

Development and Differentiation (continued)


GRIFIN Rat 50 MouseIgG1 IF,WB Purified 50µg 611502


HERC2 Ms 17 MouseIgG1 WB Purified 50µg 612366

Jagged1 Hu 21 MouseIgG1 WB Purified 50µg 612346

Janusin Hu,Ms,Rat 9 MouseIgG1 IF,IP,WB Purified 50µg 610629

KNP-1 (HES1) Chick,Dog,Hu,Ms,Rat 35 MouseIgG1 IF,WB Purified 50µg 611596

150µg 611597

Lamin A/C Chick,Dog,Hu,Ms,Rat 14 MouseIgG1 IF,WB Purified 50µg 612162


LAR Bov,Hu,Ms,Rat 7 MouseIgG2a IF,IP,WB Purified 50µg 610350


LR11 Hu,Ms,Rat 48 MouseIgG2a IF,WB Purified 50µg 611860


Dros 48 MouseIgG2a IF,IHC,WB Purified 50µg 612633

MASH1 Ms,Rat 24B7.2D11 MouseIgG1 WB Purified 100µg 556604

M-Cadherin Ms,Rat 5 MouseIgG2a IF,WB Purified 50µg 611100

150µg 611101

Mena Chick,Hu,Ms,Rat 21 MouseIgA IF,WB Purified 50µg 610692

150µg 610693

Myelin Basic Protein Gt,Ms,Prim,Rab,Rat,Sheep MyelinBP MouseIgG2b WB Purified 100µg 559904

N-Cadherin Chick,Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 50µg 610920

150µg 610921

Neurogenin 3 Hu,Ms,Rat 7 MouseIgG1 IF,IP,WB Purified 50µg 610790

Neuroglycan C Rat 1 MouseIgG1 IF,WB Purified 50µg 610986

Neuropilin-2 Rat 54 MouseIgG1 IF,WB Purified 50µg 611262

Nogo-A Rat 17 MouseIgG1 IF,WB Purified 50µg 612238

Pax-5 Hu,Ms 24 MouseIgG1 IF,WB Purified 50µg 610862

150µg 610863

Pit-1 Rat 29 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610223

R-Cadherin Ms,Rat 48 MouseIgG1 IP,WB Purified 50µg 610414

150µg 610415

Sema4C Hu 37 MouseIgG1 WB Purified 50µg 612486

TOAD-64 Ms,Rat Polyclonal RabbitIg WB Purified 100µL 551357

Tubby Ms 40 MouseIgG1 WB Purified 50µg 612106

V-1 Hu,Ms,Rat 49 MouseIgG2b IF,WB Purified 50µg 611830


ZPR1 Ms 8 MouseIgG1 WB Purified 150µg 612129




Growth Factor Related


B2 Bradykinin Receptor Chick,Dog,Hu,Ms,Rat 20 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610451

MouseIgG2b IF,IHC,IP,WB Purified 150µg 610452

c-erbB-2 Hu 42 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610161

150µg 612162

Endopeptidase 3.4.24.16 Hu,Ms,Rat 35 MouseIgA WB Purified 50µg 611240


Glucocorticoid Receptor Hu 41 MouseIgG1 IF,WB Purified 50µg 611226


IL-6 Hu MQ2-13A5 RatIgG1 FCM,IC Purified 100µg 554542

Neu NA/LE 500µg 554541

LEDGF Hu 26 MouseIgG1 IF,WB Purified 50µg 611714


Neurotensin Receptor 3 Hu 48 MouseIgG1 IF,WB Purified 50µg 612100


Thrombin Receptor (PAR1) Hu,Ms 14 MouseIgG1 IF,WB Purified 50µg 611522

150µg 611523

TrkB Ms,Rat 47 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610101


Polyclonal RabbitIg IF,WB Purified 50µg 611640


VEGF Hu G153-694 MouseIgG2b IF,IHC(F) Purified 500µg 555036

G143-850 MouseIgG2a IP,WB Purified 500µg 554539

A principal class of intercellular signaling molecules is the protein

growth factors. Neurotrophins and the neurotrophin receptors

represent a family of molecules that regulate growth and survival in

the nervous system. The first functional neurotrophin described is

nerve growth factor (NGF). NGF was discovered 50 years ago as a

molecule that promoted the survival and differentiation of sensory

and sympathetic neurons. NGF also affects additional processes such

as differentiation and microtubule assembly in some cells. Other

neurotrophin family members include brain-derived neurotrophic

factor (BDNF), neurotrophin-3 (NT3), and NT4/5. Neurotrophins have

specific patterns of expression and affect different and overlapping

neuronal populations. Other molecules that have been shown to

play roles in proliferation are GDNF (glial cell line derived neuro-

trophic factor), CNTF (ciliary neurotrophic factor), TGF-b, LIF, IL-6,

VEGF, and FGF. Different growth factors stimulate different and over-

lapping cell types in the nervous system. TGF-b is related to injury

response and produced by both glia and neurons. VEGF appears to

have neuro-protective properties.

Soluble and membrane embedded neurotrophic factors bind to

specific receptors on responsive neurons. Neurotrophins bind the

Trk family of tyrosine kinase receptors (TrkA, TrkB, TrkC) and the

p75 neurotrophin receptor (p75NTR, CD271), the low affinity NFG

receptor subunit. The Trk receptors are responsible for most of the

survival and growth properties of the neurotrophins. The actions of

p75NTR fall into two categories. First, p75NTR is a Trk co-receptor

that can enhance or suppress neurotrophin-mediated activity.

Second, p75NTR can act autonomously in apoptosis or cell survival.

TrkB binds the neurotrophins NT3, NT4/5, and BDNF. Additionally,

CNTF and LIF act as growth factors and signal through gp130 and

the JAK/STAT pathway.

Growth Factor Related



Ion Channels and Transporters


Aquaporin 2 Ms,Rat Polyclonal RabbitIg WB Purified 200µL 550648

Ca 2+ Channel Ca v1.2 Ms,Rat Polyclonal RabbitIg WB Purified 50µL 550716

IHC,WB Purified 200µL 550715

Connexin-43 Chick,Dog,Hu,Ms,Rat 2 MouseIgG1 IF,WB Purified 50µg 610061

150µg 610062

Erg2 Ms,Rat 46 MouseIgG1 IF,WB Purified 50µg 611076

GTRAP48 Rat 20 MouseIgG1 WB Purified 50µg 612672


K+ Channel a Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 50µg 611248


K+ Channel SK3 Hu,Rat Polyclonal RabbitIg WB Purified 200µL 550654

Na+, K+ ATPase b 2 Hu,Ms,Rat 35 MouseIgG2b IF,WB Purified 50µg 610914


Na+, K+ ATPase b 3 Hu 46 MouseIgG1 IF,WB Purified 50µg 610992


NHE-1 Dog,Hu,Ms,Rat 54 MouseIgG1 IF,WB Purified 50µg 611774

150µg 611775

PCMT-II Bov,Dog,Hu 4 MouseIgG1 IF,IP,WB Purified 50µg 610772

PMCA2 Hu,Ms,Rat 28 MouseIgG1 IF,WB Purified 50µg 610932

Transporting molecules across a membrane can occur as either

passive transport or active transport. Passive transport does not

utilize extra energy to move molecules across the cell membrane.

Coupled transport pairs the movement of one molecule down its

gradient to move another molecule against a gradient. Examples of

passive transport are: • Gated (ligand-gated Na+ channel)

• Non-gated ion channels (K+ leak channel)

• Carriers transporters:

– Uniporter (GLUT1 uniporter)

– Symporter (Na+/glucose symporter)

– Antiporter (Na+/Ca2+ antiporter)

Transporters utilize either an electrochemical or a concentration

gradient as a source of energy. Molecules get transported down the

gradient or against the gradient for coupled transport pairs.

Active transport requires ATP to drive unfavorable movements of

molecules against their electrochemical gradient.

Examples of active transporter are Na+/K+ ATPase and ABC

transporters (such as MDR-1). They are ATP-powered pumps that

use ATP hydrolysis as an energy source, transporting against the

potential gradient. Membrane transport proteins play an important

role in establishing the membrane potential and producing action

potentials.

The membrane potential is the balance of charges between the

inside and outside of the cell and is not equal to zero. The mamma-

lian cell typically has a membrane potential of –70 millivolts inside

the cell. This negative membrane potential is due to three contribu-

tors of negative charges inside the cell: 1) K+ leak channels let

positive cations out, 2) Na+/K+ ATPase pumps three Na+ ions out of

the cell and only returns two K+ ions into the cell, and 3) protein

anions inside the cell have negative charge.

The action potential is a change in this resting potential, which

results as a way to communicate electrically within nerve cells.

The action potential is the electrical signal, but a chemical

signal exists between nerve cells. This takes place in the form of

neurotransmitters. As the action potential propagates down the

nerve axon, it depolarizes the cell membrane and opens voltage-

gated calcium channels. Calcium enters the cell at the nerve terminal

to cause vesicles to fuse with the plasma membrane and release

neurotransmitters. Once neurotransmitters enter the synaptic cleft,

calcium binds ligand-gated Na+ channels at the postsynaptic cell and

opens them. Na+ enters the postsynaptic cell to start another action

potential.

Ion Channels and Transporters




Neurotransmitter Receptor Related


Acetylcholine Receptor a Ms,Rat 26 MouseIgG2a IF,WB Purified 50µg 610988


Acetylcholine Receptor b Hu,Rat 74 MouseIgG2b IF,IP,WB Purified 50µg 610283


APM Hu,Rat 12 MouseIgG2a IF,WB Purified 50µg 611166


b Arrestin1 Hu,Ms,Rat 10 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610550


Carbonic Anhydrase XIV Ms,Rat 49 MouseIgG2a WB Purified 50µg 612140

COMT Ms,Rat 4 MouseIgG1 WB Purified 50µg 611970

EAAT2 Rat 20 MouseIgG1 IF,WB Purified 50µg 611654

GABA BR2 Ms,Rat 1 MouseIgG1 IF,WB Purified 50µg 611744

GAD65 Hu,Ms,Rat,Pig GAD-6 MouseIgG2a,k IHC(Fr),IP,WB Purified 100µg 559931

GAD67 Rat 2 MouseIgG2a IF,WB Purified 50µg 611604

GluR d 2 Ms,Rab,Rat 48 MouseIgG1 IF,WB Purified 50µg 611006

GluR2 Dog,Monk,Rat 6C4 MouseIgG2a IHC(F),RIA,WB Purified 100µg 556341

GluR2/4 Hu,Monk,Rat 3A11 MouseIgG2a IHC(Fr),IP,WB Purified 100µg 556307

GluR5/6/7 (N-Terminal) Hu,Monk,Rat 4F5 MouseIgM EM,IF,IHC Ascites 250µL 556306

Glutamine Synthetase Hu,Rat 6 MouseIgG2a IF,IHC,IP,WB Purified 50µg 610517


mGluR1 Ms,Rat 20 MouseIgG1 IF,WB Purified 50µg 610964


mGluR1a Rat G209-488 MouseIgG1 IHC(Fr),WB Purified 100µg 556331

G209-2048 MouseIgG1 IHC(Fr) Purified 100µg 556389

NMDAR1 Hu,Monk,Rat 54.1 MouseIgG2a EM,IHC,WB Purified 100µg 556308

NMDAR2A Ms,Rat 5 MouseIgG1 WB Purified 50µg 612286

NMDAR2B Hu,Ms,Rat 13 MouseIgG2b IF,IHC,IP,WB Purified 50µg 610416


Purinergic Receptor P2X7 Rat Polyclonal RabbitIg WB Purified 50µL 550694

WB Purified 200µL 550693

Serine Racemase Ms,Rat 29 MouseIgG1 IF,WB Purified 50µg 612052


Serotonin Receptor 5-HT 2AR Hu,Monk,Rat G186-1117 MouseIgG1 IHC(Fr),WB Purified 100µg 556326

Serotonin Receptor 5-HT 2BR Hu,Rat A72-1 MouseIgG1 IHC(Fr),WB Purified 100µg 556334

Serotonin Receptor 5-HT 2CR Hu,Rat A4-2 MouseIgG1 IHC(Fr),WB Purified 100µg 556335

Substance P Pig,Rat NC1/34 RatIgG2a EM,IHC(Fr),RIA Purified 100µg 556312

Neurotransmitters are the chemical signals that relay electrical

signals between a neuron and another cell, and thus mediate many

responses within the organism. Hundreds of small molecular weight

molecules have been described as neurotransmitters. They can be

amino acids or monoamines, such as L-glutamate, GABA, dopamine,

or acetylcholine and peptides such as members of the opioid family

or hormones. Once the molecules of a neurotransmitter are released

from the synaptic vesicles of the cell, they bind to specific recep-

tors on the surface of the postsynaptic cell. Ligand-gated ionotropic

receptors are represented by receptors for acetylcholine, GABA, and

glutamate and are made up of several subunits. The metabotropic

receptors such as norepinephrine, serotonin, dopamine, and other

neuropeptides are single, transmembrane proteins that function

through GTP-binding proteins. Nerve transmissions that terminate

at a muscle cell, at the neuromuscular junction, are mediated by

acetylcholine receptors. The GABA receptor mediates the action of

g-aminobutyric acid, a major inhibitor transmitter in the brain and

spinal cord. A majority of synapses in the central nervous system

utilize glutamate as a neurotransmitter. The ionotropic receptors

glutamate receptors are subdivided into two types – receptors for

N-methyl D-aspartate (NMDAR) and non-NMDA receptors for AMPA

and kainate.

Neurotransmitter Receptor Related



Pathology


ABR Hu,Ms,Rat 24 MouseIgG1 IF,WB Purified 50µg 611122


a-Synuclein Hu,Ms,Rat 42 MouseIgG1 IF,IP,WB Purified 50µg 610786


ApoE Dog,Hu,Ms,Rat 32 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610449


APP-BP1 Dog,Hu,Ms,Rat 20 MouseIgG2a IF,WB Purified 50µg 611864


Ataxin-2 Hu,Rat 22 MouseIgG1 IF,WB Purified 50µg 611378

b-Amyloid (a.a. 1-42) Hu Polyclonal RabbitIg Blot,ELISA, Purified 25µg 556501 IHC(Fr),RIA

b-Synuclein Rat 8 MouseIgG1 WB Purified 50µg 612508

Ceruloplasmin Hu,Ms,Rat 8 MouseIgG1 IF,WB Purified 50µg 611488


Choline Acetyltransferase Rat Polyclonal GoatIg IHC,WB Purified 100µL 552623

Cox-2 Chick,Hu,Ms 33 MouseIgG1 WB Biotin 50µg 610205

WB Biotin 150µg 610206



DDX1 Hu 22 MouseIgG1 IF,WB Purified 50µg 611206

150µg 611207

ERAB Bov,Dog,Hu,Ms,Rat 23 MouseIgG1 IF,WB Purified 50µg 611086


Fascin Ms 5 MouseIgG2b WB Purified 50µg 612608


HAP1 Ms,Rat 1 MouseIgG1 IF,WB Purified 50µg 611302

HDJ-2 Hu 30 MouseIgG1 WB Purified 150µg 611873

HIP1 Hu,Ms,Rat 13 MouseIgG1 WB Purified 50µg 612314


Chick,Dog,Hu,Ms,Rat 34 MouseIgG2a WB Purified 50µg 612316

Hip1R Ms 44 MouseIgG1 WB Purified 50µg 612118

MCM Dog,Hu,Ms,Rat 24 MouseIgG1 WB Purified 150µg 611772

150µg 611773

MnSOD Dog,Hu,Ms,Rat 19 MouseIgG1 WB Purified 50µg 611580

150µg 611581

Neuronal Pentraxin Hu,Rat 22 MouseIgG1 IF,IP,WB Purified 50µg 610369


Nicastrin Chick,Dog,Hu,Ms,Rat 35 MouseIgG2a WB Purified 50µg 612290

OPA1 Hu 18 MouseIgG1 IF,WB Purified 50µg 612606


PEX1 Chick,Dog,Hu,Ms,Rat 1 MouseIgG1 WB Purified 50µg 611718


PEX19 Hu 32 MouseIgG1 IF,WB Purified 50µg 611598

Neurotransmitter Receptor Related (continued)


Tyrosine Hydroxylase Ms,Rat 45 MouseIgG1 WB Purified 50µg 612300

Rat TOHA1 MouseIgG1 EM,IHC(Fr) Purified 100µg 556311

VAChT Ms,Rat null Goat IHC(Fr) Serum 50µL 556337




Pathology (continued)


SIP1 Dros 4 MouseIgG1 IF,IHC,WB Purified 50µg 612655

Hu 4 MouseIgG1 IF,WB Purified 50µg 611256

SMN Hu 8 MouseIgG1 IF,IHC,WB FITC 50µg 612586

IF,IHC,WB FITC 150µg 612587

Dog,Hu,Ms,Rat 8 MouseIgG1 IF,IHC,WB Purified 50µg 610646


TCBP-49 Ms,Rat 30 MouseIgG2a WB Purified 50µg 612178


UBE3A Hu,Ms 13 MouseIgG1 IF,WB Purified 50µg 611416

Synaptic Vesicles


a-Methylacyl-CoA Racemase (AMACR) Hu,Ms,Rat 15 MouseIgG1 WB Purified 50µg 612082


Adaptin a Dog,Hu,Ms,Rat 8 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610501


Adaptin b Chick,Dog,Frog,Hu,Ms,Rat 74 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610381


Adaptin d Hu 18 MouseIgG1 IF,WB Purified 50µg 611328


Adaptin e Hu null IgG1 IF,WB Purified 50µg 612018


Adaptin g Dog,Hu,Ms,Rat 88 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610385


AF6 Hu,Ms,Rat 35 MouseIgG2a IF,WB Purified 50µg 610732

150µg 610733

a-/b -SNAP Chick,Hu,Ms,Rat 32 MouseIgG2a IF,WB Purified 50µg 611898

Amphiphysin Ms,Rat 15 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610714

AP180 Hu,Ms,Rat 34 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610469


ARF3 Chick,Dog,Frog,Hu,Ms,Rat 41 MouseIgG1 IP,IF,WB Purified 50µg 610784

b-NAP Ms,Rat 18 MouseIgG1 IF,WB Purified 50µg 610892


BRAMP2 Ms,Rat 54 MouseIgG1 IF,IP,WB Purified 50µg 610856


CDCrel-1 Hu 49 MouseIgG1 IF,WB Purified 50µg 612386

Cellugyrin Ms,Rat 24 MouseIgG1 IF,WB Purified 50µg 611128

Nerves transmit signals from one to another at contact points called

synapses via a process called synaptic transmission. The two types of

synaptic transmission are electrical and chemical. Electrical transmis-

sion occurs at gap junctions. Chemical synaptic transmission occurs

at synaptic vesicles via neurotransmitters released at the presynaptic

neuron that diffuse across the synaptic cleft and are detected by

receptors on the postsynaptic cell. The postsynaptic cell can be a

neuron, a muscle, or a secretory or gland cell.

Synaptic vesicles are organelles situated at the distal terminus of the

presynaptic neuron. In the typical sequence of events, the action

potential arrives at the axon terminal and triggers influx of calcium

ions that signal exocytosis, the fusion of synaptic vesicles with the

cell membrane. SNARE complex proteins then regulate membrane

fusion. SNARE complex consists of the SNAPs (soluble NSF attach-

ment proteins), NSF (N-ethyl-maleimide-sensitive factor) along with

the receptor proteins (SNAREs such as synaptobrevin, synapto-

tagmin, syntaxin, and SNAP-25). Several Rab proteins expressed in

neuronal cells and are associated with synaptic vesicles.

Synaptic Vesicles and Post-Synaptic Proteins



Synaptic Vesicles (continued)


Chromogranin A Rat 5 MouseIgG1 IF,WB Purified 50µg 611844

Chromogranin B Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 50µg 611646


Clathrin Heavy Chain Chick,Dog,Hu,Ms,Rat 23 MouseIgG1 IF FITC 50µg 611784



Complexin 2 Dog,Rat 48 MouseIgG1 IF,IP,WB Purified 50µg 610728


CSP Rat 16 MouseIgG1 IF,WB Purified 50µg 612662


DLP1 Dog,Hu,Ms,Rat 8 MouseIgG1 IF,WB Purified 50µg 611112

150µg 611113

Dynamin Chick,Dog,Hu,Ms,Rat 41 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610245


Dynamin II Hu,Ms,Rat 27 MouseIgG2a IF,WB Purified 50µg 610263

150µg 610264

GS15 Bov,Dog,Frog,Hu,Ms,Rat 19 MouseIgG1 IF,WB Purified 50µg 610960


GS27 Dog,Hu 25 MouseIgG1 IF,WB Purified 50µg 611034


GS28 Hu,Ms,Rat 1 MouseIgG2a IF,WB Purified 50µg 611184


Mint1 Ms,Rab,Rat 23 MouseIgG1 IF,WB Purified 50µg 611028


Mint2 Ms,Rat 18 MouseIgG1 IF,WB Purified 50µg 611032

Mint3 Hu,Ms,Rat 32 MouseIgG1 IF,WB Purified 50µg 611380

Munc-18 Rat 31 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610336


Neurexin I Dog,Hu,Ms,Rat 17 MouseIgG1 IF,WB Purified 50µg 611882

NSF Chick,Dog,Hu,Ms,Rat 7 MouseIgG2b IF,WB Purified 50µg 612272


p47A Dog,Hu,Ms,Rat 26 MouseIgG1 IF,WB Purified 50µg 610890



panMunc13 Rat 32 MouseIgG1 IF,WB Purified 50µg 610998


ProSAP1 Rat 2 MouseIgG1 WB Purified 50µg 612274

Rab11 Chick,Dog,Hu,Ms,Rat 47 MouseIgG2a IF,IHC,WB Purified 50µg 610656

150µg 610657

Rab3 Dog,Rat 9 MouseIgG2a IF,IHC,IP,WB Purified 50µg 610379

Rab5 Hu 1 MouseIgG2a IF,IHC,WB Purified 50µg 610724

150µg 610725

Rim Ms,Rat 26 MouseIgG1 IF,WB Purified 50µg 610906


Sec8 Chick,Dog,Hu,Ms,Rat 14 MouseIgG2b IF,IP,WB Purified 50µg 610658

150µg 610659

SNAP-25 Ms,Rat 20 MouseIgG1 IF,IP,WB Purified 50µg 610366





Post-Synaptic Proteins


Arc Rat 40 MouseIgG1 WB Purified 50µg 612602


Gephyrin Chick,Hu,Ms,Rat 45 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610584


Dros 45 MouseIgG1 IF,IHC,IP,WB Purified 50µg 612632

GRIP Rat 32 MouseIgG1 IF,WB Purified 50µg 611318


MUPP1 Ms,Rat 43 MouseIgG1 IF,WB Purified 50µg 611558


Neurabin Ms,Rat 52 MouseIgG1 IF,WB Purified 50µg 611088

Dros 52 MouseIgG1 IF,WB Purified 50µg 612639

Neurabin II Rat 17 MouseIgG1 WB Purified 50µg 612166

PSD-95 Ms,Rat 16 MouseIgG1 IF,IP,WB Purified 50µg 610495


SCAMP1 Hu 22 MouseIgG1 WB Purified 150µg 612087

Vesl-1L Rat 8 MouseIgG1 IF,WB Purified 50µg 611174

Synaptic Vesicles (continued)


Synapsin I Ms,Rat 8 MouseIgG1 IF,WB Purified 50µg 611392


Synapsin IIa Hu,Rat 1 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610666

Synaptogyrin Rat 6 MouseIgG1 IF,IHC,IP,WB Purified 50µg 610597

Synaptophysin Ms,Rat 2 MouseIgG1 WB Purified 50µg 611880

Synaptotagmin Hu,Rat 41 MouseIgG1 IF,IP,WB Purified 50µg 610433


Synaptotagmin V Ms,Rat 46 MouseIgG1 WB Purified 50µg 612284

Syncollin Rat 17 MouseIgG2b WB Purified 50µg 612468

Syndapin I Ms,Rat 32 MouseIgG1 WB Purified 50µg 611810


Syntaphilin Ms,Rat 30 MouseIgG1 WB Purified 50µg 612324

Syntaxin 11 Hu,Ms 32 MouseIgG1 IF,WB Purified 50µg 611282


Syntaxin 4 Dog,Hu,Ms,Rat 49 MouseIgG1 IF,IP,WB Purified 50µg 610439


Syntaxin 6 Chick,Dog,Frog,Hu,Ms,Rat 30 MouseIgG1 IF,IP,WB Purified 50µg 610635


Syntaxin 8 Dog,Hu,Ms,Rat 48 MouseIgG2a IF,WB Purified 50µg 611352


Tomosyn Hu,Ms,Rat 15 MouseIgG1 IF,WB Purified 50µg 611296




For Probenecid Sensitive Applications

description size cat.no.

BD PBX Calcium Assay Kit 10plates 640175

100plates 640177

For Standard Applications


BD Calcium Assay Kit 10plates 640176

100plates 640178

Assay Kits and Reagents for Bioimaging and Flow Cytometry

BD introduces two new calcium assay kits for detecting calcium flux

in living cells. The proprietary formulations combine a non-fluores-

cent calcium indicator that becomes activated inside the cell, along

with a signal enhancer to provide an easy-to-use, no-wash assay

format with increased signal to background. In most cases, the dye

kits perform better than kits currently available on the market with

no known interference to ligands (small molecules, peptide/proteins,

and lipids). Also, the “addition artifact” commonly seen with some

kits is significantly reduced with these new dye formulations.

• No-wash homogeneous assay format

• Higher signal over background compared to competitive kits

• PBX kit permits assays with reduced or no probenecid

• Ideal for all fluorescence-based calcium flux assays

The kits have been successfully used with a variety of cell lines

including CHO, HEK293, Jurkat, Hela, and U2OS.

Kits are available in two sizes providing sufficient material for either

10 plates (96, 384, or 1536 wells) or 100 plates. Additionally, for

higher volume applications, custom packaged materials can also be

provided. Please contact customer support for more details.

Calcium Flux Assays




BD Biosciences has always provided the most reliable and complete

multiplex bead array system. We have taken the performance you

trust and the ease of use you rely on and have made them even

more flexible. In addition to your need for a flexible assay system,

we also recognize the value of your precious samples and the

importance of data reproducibility, so we’ve engineered the new

BD™ Cytometric Bead Array (CBA) Flex Set system to maximize

both. Further, BD CBA Flex Sets can be used with any dual-laser

flow cytometer, or you can increase throughput by automating your

acquisition and analysis on a BD FACSArray™ bioanalyzer.

With the BD CBA Flex Set System You Can:

• Configure your multiplex assay on demand

• Get multiple results from a single, small-volume sample

• Achieve quantitative results in less time

• Use with any dual-laser flow cytometer (non stream-in-air)

• Automate sample acquisition and increase throughput with the

BD™ HTS option coupled with BD™ flow cytometers or the

plate-based BD FACSArray bioanalyzer

The new BD CBA Flex Set system is an open and configurable system

designed to be the easiest method of creating multiplex assays.

With our proprietary

conjugation chemistry, pair

optimization strategies,

and direct PE detection

reagents, you can be

assured consistent and

superior assay performance

in complex biological

samples. Each antibody

pair we develop is

evaluated for dynamic

range, sensitivity, and

parallel titration to native

biological samples. We have also specifically formulated the assay

diluent and wash buffers for each assay type to reduce potentially

detrimental effects on assay performance when using serum and

plasma samples. By avoiding the streptavidin-biotin-PE detection

method employed by other assays, our direct PE detection reagents

minimize the risk of increased background often caused by

endogenous biotin in serum and lysate samples. Thus, BD CBA Flex

Sets provide a reliable and flexible method for quantitative detection

of multiple analytes in a single serum, plasma, tissue culture

supernatant, or cell lysate sample.

102 10 103 4 105

Red-A

2

3

4

10

10

10

10

5

NIR

-A

BD Cytometric Bead Array Flex Set Reagents

description react bead size cat.no.

Btk (Y551) Hu D5 100tests 558236

eNOS (S1177) Hu C7 100tests 558239

ERK1/2 (T202/Y204) Hu,Ms,Rat C4 100tests 558234

Itk (Y511) Hu,Ms C6 100tests 558230

JNK1/2 (T183/Y185) Hu,Ms,Rat B5 100tests 558235

p38 (T180/Y182) Hu,Ms,Rat B6 100tests 558233

PLC-g (Y783) Hu,Ms,Rat B7 100tests 558228

RSK (T573) Hu,Ms,Rat D7 100tests 558240

Stat1 (Y701) Hu,Ms C5 100tests 558222

Syk (Y352) Hu,Ms,Rat B9 100tests 558237

ZAP-70 (Y319) Hu,Ms B8 100tests 558229

Stat1 Hu,Ms,Rat D4 100tests 558227

Syk Hu,Ms,Rat B9 100tests 558239

ZAP-70 Hu,Ms,Rat B8 100tests 558232

Cell Signaling Master Buffer Kit 100tests 558223

500tests 558224

BD Cytometric Bead Array Flex Set Reagents

Visit bdbiosciences .com/flexset for complete product listings.



High-Content Bioimaging Certified Antibodies


53BP1 19 IF,WB Purified 50µg 612522


b-Tubulin 5H1 ICC,WB Purified 100µg 556321

Actin C4 IF,WB Purified 50µg 612656


ATP Synthase b 10 IF,WB Purified 50µg 612518


Bcl-x 44 IF,IHC,IP,WB Purified 50µg 610746


Bid 7 IF,WB Purified 50µg 611528


Bub3 31 IF,WB Purified 50µg 611730


c-IAP-1 B75-1 IP,WB Purified 100µg 556533

c-Jun (pS63) 2 IHC(F),WB Purified 100µg 558036

Cab45 30 IF,WB Purified 50µg 611064


Calnexin 37 IF,IHC,IP,WB Purified 50µg 610523


Calretinin 34 IF,WB Purified 50µg 610908


Caspase-2 G310-1248 WB Purified 50µg 551093


Caspase-3 19 IF,IHC,IP,WB Purified 50µg 610322


Caspase-3, Active Form C92-605 FCM,IHC(Fr),IP Purified 25µg 559565

Caspase-6 B93-4 WB Purified 100µg 556581

Caspase-7 8-1-47 IP,WB Purified 50µg 551236

IP,WB Purified 150µg 551237

Caspase-8 Polyclonal WB Serum 100µL 551234

Caspase-14 32 IF,WB Purified 50µg 611510


Caveolin 1 C060 FCM,IF,IHC,IP,WB Purified 50µg 610057

FCM,IF,IHC,IP,WB Purified 150µg 610058

Cdc25B 23 IF,IHC,IP,WB Purified 50µg 610527


Cdc25C C2-2 WB Purified 100µg 556576

CDC5L 21 IF,WB Purified 50µg 612362

BD Biosciences is now offering a rapidly growing collection of

antibody reagents that have been tested and certified for research

applications in automated bioimaging (high-content screening, HCS)

and fluorescence microscopy.

These Bioimaging Certified reagents are optimal for use in

high-content applications when reproducibility is essential.

BD Biosciences bioimaging reagents are certified for optimal

performance based on an internal test program. For BD to certify

these reagents, each antibody must pass a number of tests that

focus on high content applications.

Theses tests include:

• Exceeding a signal intensity threshold over background in

commonly used cell lines

• Working with widely used fixation and permeabilization

methods

• Localizing to the appropriate region within a cell

• Showing an appropriate response (ie, redistribution) following

stimulation

Visit bdbiosciences .com/bcr for the latest product releases.




High-Content Bioimaging Certified Antibodies (continued)


Cdk2 55 IF,IHC,IP,WB Purified 50µg 610145


Cdk4 DCS-35 IF,IP,WB Purified 100µg 559693

Cdk7 MO-1 IF,IHC(Fr),IP,WB Purified 100µg 556345

Cdk8 F9-1075 WB Purified 50µg 552056


CGBP 35 IF,WB Purified 50µg 612334


Chk2 19 IF,WB Purified 50µg 611570


Crk 22 IF,IHC,IP,WB Purified 50µg 610035


Cyclin A 25 IF,WB Purified 50µg 611268


Cyclin B1 GNS-11 FCM,IF,IP,WB Purified 100µg 554178

FCM,IF,IP,WB Purified 250µg 554179

Cyclin B1 (a.a 1-21) GNS-1 FCM,IF,IHC(F), Purified 100µg 554176 IHC(Fr),IP,WB

FCM,IF,IHC(F), Purified 250µg 554177 IHC(Fr),IP,WB

Cyclin E HE12 FCM,WB Purified 50µg 551159

FCM,WB Purified 150µg 551160

Cytochrome c 6H2.B4 IP Purified 100µg 556432

D4-GDI Polyclonal WB Serum 100µL 556498

eIF-4E 87 IF,IP,WB Purified 50µg 610269


IF,IP FITC 50µg 611624

IF,IP FITC 150µg 611625

eIF-5a 26 WB Purified 50µg 611976


eNOS (pS1177), Phospho-Specific 19 FCM,WB Purified 50µg 612392

FCM,WB Purified 150µg 612393

eNOS (pS633) 37 WB Purified 50µg 612664


eNOS/NOS Type III 3 IF,IHC,IP,WB Purified 50µg 610296


33 IF,IP,WB Purified 50µg 610427


ERK (pan) 16 IF,IHC,IP,WB Purified 50µg 610123


ERK1/2 (pT202/pY204) 20A WB Purified 50µg 612358


ERK3 30 IF,IHC,IP,WB Purified 50µg 610105


Ezrin 18 IF,IHC,IP,WB Purified 50µg 610602


FADD 1 IF,IP,WB Purified 50µg 610399






FAK 77/FAK IF,IHC,IP,WB Purified 50µg 610087


FAK (pY397) 14 IF,WB Purified 50µg 611722


18 IF,WB Purified 50µg 611806


Fatty Acid Synthase 23 IF,WB Purified 50µg 610962


FEN-1 21 IF,WB Purified 50µg 611294


FIN13 7 IF,WB Purified 50µg 610910


Fnk 53 IF,WB Purified 50µg 611178


Fractin Polyclonal IHC,WB Purified 100µL 551527

GRB2 81 IF,IHC,IP,WB Purified 50µg 610111


GSK-3b 7 IF,IHC,IP,WB Purified 50µg 610201


GSK-3b (pY216) 13a WB Purified 50µg 612312


Hsp60 24 IF,WB Purified 50µg 611562

IF FITC 50µg 611958


IKKa B78-1 IP,WB Purified 100µg 556532

IRAK 7 IF,IP,WB Purified 50µg 610754


KAP 39 IF,IHC,IP,WB Purified 50µg 610334

Laminin B2 22 IF,IP,WB Purified 50µg 610722

MCM6 1 IF,WB Purified 50µg 611622


MDM2 SMP14 IF,IHC(F),IHC(Fr), Purified 100µg 556353 IP,WB

MEK1 25 IF,IHC,IP,WB Purified 50µg 610121


Metablastin 53 IF,WB Purified 50µg 611146


NF-kB p65 20 IF,WB Purified 50µg 610868


p19Skp1 52 IF,IHC,IP,WB Purified 50µg 610530

p23 16 WB Purified 50µg 612320


p27Kip1 G173-524 IP,WB Purified 100µg 554069

p32 20 IF,WB Purified 50µg 612396


p38 MAPK (pT180/pY182) 36 IF,WB Purified 50µg 612288


p47A 26 IF,WB Purified 50µg 610890







p53 DO-1 IHC(F),IHC(Fr), Purified 100µg 554293 IP,WB

p54nrb 3 IF,WB Purified 50µg 611278


p55Cdc 41 IF,WB Purified 50µg 611242


PARP 4C10-5 Blot,FCM,IP,WB Purified 100µg 556494

PARP, Cleaved Form F21-852 FCM,IP,WB Purified 50µg 552596

FCM,IP,WB Purified 150µg 552597

Paxillin 177 IF,IP,WB Purified 50µg 610568


PCNA PC10 IC/FCM,IHC(F), Purified 100µg 555566 IHC(Fr)

IC/FCM Biotin 100µg 555567

Phospholipase Cg1 10 IF,IHC,IP,WB Purified 50µg 610027


PI3-Kinase 4 IF,IHC,IP,WB Purified 50µg 610045


PI3-Kinase C2b 22 IF,WB Purified 50µg 611342


PI4-Kinase b 7 IF,WB Purified 50µg 611816


PKARI 18 IF,IHC,IP,WB Purified 50µg 610165




PKARIIa 40 IF,IP,WB Purified 50µg 612242


Plakophilin 2 28 IF,IP,WB Purified 50µg 610788


PNUTS 47 IF,WB Purified 50µg 611060


Poly ADP-Ribose 10H IF Purified 50µg 550781

Psme3 47 IF,WB Purified 50µg 611180


PTRF 4 IF,WB Purified 50µg 611258


Rac1 102 IF FITC 50µg 610652


RACK1 20 IF,IHC,IP,WB Purified 50µg 610177


Ran 20 IF,IHC,IP,WB Purified 50µg 610340


RanBP3 52 IF,WB Purified 50µg 612050


RAP30 15 IF,WB Purified 50µg 612176


Rb (a.a. 300-380) G3-349 IP,WB Purified 100µg 554140

Rb (a.a. 332-344) G3-245 FCM,IF, Purified 100µg 554136 IC/FCMBlock,IHC(F), IHC(Fr),IP,GS,WB





S100P 16 IF,IHC,IP,WB Purified 50µg 610306


Sec31A 32 IF,WB Purified 50µg 612350


SII 7 IF,WB Purified 50µg 611204


Smac/DIABLO 7 IF,WB Purified 50µg 612244


Sos1 25 IF,IHC,IP,WB Purified 50µg 610095


SRC-1 8 IF,WB Purified 50µg 612378

Stat1 42 IF,IHC,IP,WB Purified 50µg 610185


Stat1 (pY701) 14 FCM,IF,IP,WB Purified 50µg 612132


4a FCM,IF,IP,WB Purified 50µg 612232


Stat3 (pS727) 49 IF,WB Purified 50µg 612542


Stat3 (pY705) 4 WB Purified 50µg 612356


Stat6 23 IF,WB Purified 50µg 611290


Stat6 (pY641) 18 IF,WB Purified 50µg 611566


TOK-1 13 WB Purified 50µg 612326

Topo IIa 31 IF,WB Purified 50µg 611326


Topo IIb 40 IF,WB Purified 50µg 611492


Visit bdbiosciences .com/bcr for our latest product releases.




Screening and Development of Antibodies for use in Cell-Based Assays using Immunofluorescence Microscopy

IntroductionHigh content imaging is rapidly becoming a mainstay in pharmaceutical and life science research laboratories.1 The power of imaging resides in its ability to measure not only fluorescence intensity changes within cells, but also molecular rearrangements and morphological features. In recent years, the availability of a new generation of automated imaging platforms has created a unique opportunity to develop novel cell-based assays.2-4 High content microscopy has also been advanced by the abundant use of antibodies in immunofluorescence applications. These antibody reagents have enabled high resolution localization of important protein targets in individual cells. For example, antibodies can be used to detect specific stages in apoptosis, in the activation of signaling cascades, and in the cell cycle. However, many of the antibodies currently available have not been specifically tested for use in automated imaging applications. In addition, the promise of high content screening relies on the multiplexing of cellular information; the ability to do this however, is often limited when using standard unlabeled primary antibodies in combination with labeled secondary antibodies due to species cross-reactivity.

To facilitate the use of antibodies for imaging applications, BD Biosciences embarked on a screening process using automated imaging to evaluate a large library of monoclonal antibodies. The library of antibodies included specificities that recognize proteins involved in apoptosis, cancer, cell cycle, cell signaling, and neurobiology. Antibodies were tested using cell lines and methods that are relevant in automated imaging. Specific criteria regarding signal to noise, sub-cellular localization, and other important parameters were used to qualify reagents. Two distinct Bioimaging Certified Reagent product lines have resulted from this antibody screen. The first is a collection of more than 200 unlabeled primary antibodies that have been shown to have general utility in bioimaging applications. A subset of these reagents has been developed into conjugated primary antibodies in order to truly enable high content screening using antibody multiplexing. These reagents, combined with an automated imaging platform such the BD Pathway bioimager 415, 435, or 855 can enable rapid development of novel cell-based assays.

Methods for Screening of Purified AntibodiesAntibodies from the BD Biosciences library were screened for utility in bioimaging applications. The antibodies recognized proteins in different areas of biology that included apoptosis, cancer, cell cycle, cell signaling, and neurobiology. For ubiquitously expressed proteins, the antibodies were screened on three different cell lines routinely used in high content imaging applications. HeLa (human cervical carcinoma), A549 (human lung carcinoma), or U-2 OS (human osteosarcoma) cells were seeded at approximately 10,000 cells/well in 96-well clear bottom tissue culture plates (Cat. No. 353219) optimized for automated imaging applications. Eighteen hours later, cells were either treated with an appropriate compound and then fixed and permeabilized or, if not treated, immediately fixed and permeabilized.

Fixation

1. Remove culture media from wells.

2. Add 100 μL freshly prepared 3.7% formaldehyde diluted in 1x phosphate buffered saline (PBS) (Sigma F-1268) and pre-warmed to 37°C to each well.

3. Incubate at room temperature for 10 minutes.

4. Remove the formaldehyde solution and drain the residual liquid by inverting the plate briefly onto absorbent paper.

5. Wash by adding 100 μL per well of 1x PBS.

6. Proceed to either the Triton® X-100 or Methanol permeabiliza-tion steps described below.

Triton® X-100 Permeabilization

1. Remove 1x PBS and add 100 μL per well of a 0.1% Triton® X-100 solution diluted in 1x PBS.


3. Remove the 0.1% Triton® X-100 solution and drain the residual liquid.

4. Wash twice with 1x PBS.

5. Proceed to the Blocking step.

Methanol Permeabilization

1. Remove 1x PBS and add 100 μL per well of a -20°C 90% meth-anol solution diluted in water.


3. Remove the 90% methanol solution and drain the residual liquid.

4. Wash twice with 1x PBS.

5. Proceed to the Blocking step.



Technical Spotlight and Protocols



Blocking

1. Remove 1x PBS and add 100 μL per well of 1x PBS supple-mented with 3% fetal bovine serum (FBS blocking solution).


3 Proceed to the Primary Antibody Staining step.

Primary Antibody Staining

1. Remove the 3% FBS blocking solution and add 50 μL per well of primary antibody diluted to the appropriate concentration in blocking solution.

2. Incubate at room temperature for 1 hour.

3. Remove primary antibody solution.

4. Wash three times with the blocking solution.

5. Proceed with Secondary Antibody Staining.

Secondary Antibody Staining

1. Remove the 3% FBS blocking solution and add 50 μL per well of diluted secondary antibody (diluted in blocking solution). Several different second step reagents were used, including FITC goat anti-mouse IgG (Cat. No. 554001), Alexa Fluor® 555 goat anti-mouse IgG (Invitrogen), and Alexa Fluor® 488 goat anti-mouse IgG (Invitrogen). All were used at 0.25μg/well.

2. Incubate at room temperature for 1 hour in the dark.

3. Remove secondary antibody solution.

4. Wash three times with the blocking solution.

5. Add 100 μL per well of 1x PBS containing 2 μg/mL Hoechst 33342 (Sigma).

6. Cover the plate and let stand protected from light for 15 to 30 minutes.

7. Plates are now ready for imaging.

Screening of Directly Conjugated Antibodies

Essentially the same procedure as outlined above was followed, with the omission of the secondary antibody step, as the primary antibodies were directly conjugated to Alexa Fluor® dyes. After incubation with a conjugated antibody, cells were washed as described, Hoechst dye was added, and the samples were imaged.

Imaging and Analysis

Cells were imaged on a BD Pathway 855 bioimager using a 20x (0.75 NA) objective. Image analysis was done using the built in Attovision (version 1.5) software tools. Numerical data was imported into the Microsoft® Excel® based BD™ Image Data Explorer software, where cell populations were analyzed for nuclear and/or cytoplasmic fluorescence intensity.

Technical Spotlight and Protocols (continued)

Erk1/2 (pT202/pY204) During Differentiation of P19 Cells

100 101 102 103 104

Red-A

100 101 102 103 104

Red-A

100 101 102 103 104

Red-A

P19 P19+DMSO P19+Retinoic Acid

Figure 1 . Undifferentiated (left panel), DMSO differentiated (middle panel), and retinoic acid differentiated (right panel) P19 cells, were serum starved for 2 hours (orange), then treated with either PMA (green) or NGF (blue). Cells without serum starvation (red) were also included for comparison.

Erk1/2 (pT202/pY204) During Differentiation of P19 Cells

Phospho-Protein Profiling in Stem Cells

Characterization of the spatial and temporal function, or proteomic

profiling, of each component of the cellular proteome is one of the

major scientific challenges faced today. This proteomic profiling is of

critical importance in understanding how pluripotent cells develop

into an adult and complete organism, or replenish and support

existing organs. Those pluripotent cells, or stem cells, undergo a

timely coordinated gene expression inducing their differentiation.

Understanding the signaling mechanisms orchestrating those

dynamic intracellular changes is key to comprehending the biology

of stem cells. Reversible protein phosphorylation, controlled by

kinases and phosphatases, has been recognized as a universal

post-translational modification that rapidly adjusts the activity of a

given protein target. Yet despite its central role in cell biology, little

is known about the role of protein phosphorylation in stem cells and

their differentiation. Early studies demonstrated the importance of

phospho-STAT3,79 phospho-Erk, and the Src kinase in maintaining

the pluripotency of stem cells.80 Furthermore, FAK (focal-adhesion

kinase), is required for cardiogenesis of mouse embryonic stem

cells.81 Most methodologies used to monitor protein phosphorylation

in differentiating stem cells lack the ability to visually represent the

variability of this modification of individual cells and committed cell

sub-populations, thus limiting data interpretation.

BD Phosflow technology is an emerging methodology whereby the

phosphorylation of individual proteins can be monitored by flow

cytometry using highly specific antibodies as probes. The ability of

flow cytometry to identify unique cell populations opens the

possibilities for monitoring protein phosphorylation in rare cell

events types, such as T-cell precursors, developing cells, and

malignant stem cells.82,83

In the experiment described below, we have used mouse

pluripotent embryonic carcinoma stem cells (P19) to demonstrate

that BD Phosflow technology is a valuable tool for the analysis of

differentiation stages in stem cells. The P19 cell line is derived from

an embryonal carcinoma induced in a C3H/He strain mouse. The

pluripotent P19 cells can be induced to differentiate into neuronal

and glial cells in the presence of retinoic acid.84 Aggregates of P19

cells differentiate into cardiac and skeletal muscle in the presence of

dimethyl sulfoxide (DMSO). In the presence of both retinoic acid and

DMSO, cells develop as if exposed to retinoic acid only.

In Figure 1, undifferentiated, DMSO differentiated, and retinoic

acid differentiated P19 cells were serum starved for two hours. After

stimulation with either phorbol myristate acetate (PMA) or nerve

growth factor (NGF), the cells were fixed, permeabilized, stained

with Alexa Fluor® 647 conjugated anti-phospho ERK1/2

(Cat. No. 612593), and subsequently analyzed by flow cytometry.

The data shown indicates that only retinoic acid differentiated cells

show a significant increase in Erk1/2 phosphorylation upon stimula-

tion with NGF. No significant effect on Erk1/2 phosphorylation could

be detected in either retinoic acid or DMSO differentiated cells after

treatment with PMA.

Technical Spotlight





Related Immunohistochemistry Reagents


IHC Zinc Fixative 1000mL 550523

10× Zinc Fixative 500mL 552658

A ready-to-use solution, our IHC zinc fixative is an alternative to the harsh conditions of formalin fixation .

Immunohistochemistry (IHC) is a powerful technique used by

neuroscientists to identify neuron connections and to elucidate the

role of chemical messengers in relation to the architecture of the

brain. IHC with free-floating tissue sections is often impractical,

however, because it requires large amounts of antibody and

prolonged incubation periods. BD Pharmingen and

BD Transduction Laboratories antibodies, using our formalin-

free IHC Zinc Fixative (Cat. No. 550523), allow staining

to be performed on 5-μm, paraffin-embedded tissue

sections. The protocol is quick and easy to perform, the

morphologies obtained are superior to those obtained with

free-floating sections, and tissue antigenicity is retained.

In addition, our IHC antibodies are supported by a complete line of

ancillary reagents such as secondary antibodies, isotype controls, and

detection systems.

See images on the following page for phospho-specific staining of

rat brain tissues.

Visit our technical support protocol website at bdbiosciences .com

for the complete protocol on preparation and staining of

paraffin sections.

Glutamate receptor mGluR1a staining on normal rat cerebellum using our IHC Zinc Fixative: A) free-floating section and B) zinc-fixed, paraffin-embedded section. Magnification: 1× (left) and 10× (right).

A A

B B

IHC for Neuroscience Research

Technical Spotlight and Protocols (continued)



Phospho-Specific Staining of Rat Brain Tissues

ERK1 IHC images and accompanying Western blot images

ERK2 IHC image and accompanying Western blot image

JUN IHC image and accompanying Western blot image

nNos IHC image and accompanying Western blot image

Calcimeurin IHC image and accompanying Western blot image

Imaging and High Content Analysis

BD CARV II Confocal Imager

The BD CARV II confocal imager delivers high-resolution CCD

confocal imaging in an easy-to-use and cost-effective optical

package that fits on your existing microscope. High-speed multi-

point confocal scanning, combined with high quantum efficiency

CCD cameras, minimizes photobleaching and allows real-time

imaging and recording at up to 100 fps. A long life arc source

coupled to the instrument via an alignment-free light guide allows

for full spectrum (360–700 nm) confocal imaging of virtually any

fluorescent probe. Automation of internal multi-position excitation,

dichroic, and emission filter wheels permits fast multi-dimensional

imaging of up to five fluorescent probes in the same sample.

BD Pathway Bioimager

The BD Pathway bioimager is an automated, confocal, real-time,

single-cell kinetic and endpoint imaging system that has been inte-

grated into a single, compact unit. The BD Pathway bioimager was

designed to provide high-resolution, automated confocal imaging

with sophisticated imaging software to assist in developing better

cell-based assays. The system allows the user to explore biological

events without many of the restrictions of conventional microscope-

based high-content imaging systems.

The BD CARV II system provides an easy-to-use confocal upgrade to your existing microscope without sacrificing image quality or three-dimensional imaging capabilities.

The BD Pathway bioimager provides a wide range of imaging capa-bilities including kinetic, endpoint, and three-dimensional confocal imaging. The environmentally controlled imaging chamber with liquid handling provides unsurpassed live cell kinetic imaging possibilities.

Bioimaging Instrumentation





BD FACSArray Bioanalyzer

Quantitative multi-parameter cellular analysis is conducted using

flow cytometry instruments by analyzing whole cells and proteins

separately or in concert. This technique is creating new ways to

analyze neuronal cells from small sample sizes. The BD FACSArray

bioanalyzer is the primary instrument for this application.

Unlimited Possibilities

The BD FACSArray bioanalyzer from BD Biosciences, the innovation

leader in flow cytometry instrumentation, is used for fast and sensi-

tive high-content measurements of cells and proteins in cell biology,

immunology, and proteomics.

This compact flow cytometer rapidly detects and quantifies

concentrations of secreted proteins, proteins in cell lysates,

and cell-associated proteins using small sample volumes

directly from a 96-well microtiter plate through multiplexed

BD Cytometric Bead Arrays (CBAs). Furthermore, it supports

your search for answers in cellular analysis of apoptosis,

cytokine and chemokine profiling, and phosphorylation of key

signal transduction proteins. Our bioanalyzer can also be used to

assess hybridomas, cell viability, proliferation, and immunotoxicology.

For your lab, this means unlimited possibilities for both cellular

applications and multiplex bead assays.

See page 55 or visit bdbiosciences .com/flexset for a list of

BD CBA reagents.

The BD Biosciences commitment to application development is one

of our strongest distinctions in supporting customers. Carefully opti-

mized applications and availability of over 1,000 reagents support

the BD FACSArray bioanalyzer platform.

A fully integrated system, the BD FACSArray bioanalyzer is composed

of an analyzer with an automated microtiter plate sampler, a sensi-

tive optical detection subsystem, a computer workstation, intuitive

software, and validated applications. It is easy to use and requires

minimal training, so your research shows immediate success.

BD FACSArray bioanalyzer system:• Fast microtiter plate sampler

• Two-laser system

• Sensitive fluorescence detection

• Simultaneous collection of up to six parameters per particle (four fluorescence and two scatter)

• Intuitive software

• State-of-the-art digital signal processing allows up to 15,000 events per second

• Supported by BD Biosciences reagents and applications

Application Flexibility

Cellular Analysis Using Flow Cytometry

Maximize Your Productivity

The BD FACSArray bioanalyzer features an automated microtiter

plate sampler that quickly acquires from 96-well plates. The plate

sampler is controlled by flexible and easy-to-use software for imme-

diate adoption of the bioanalyzer in your laboratory. The system

software controls the instrument and sample acquisition, and is

powerful enough to support batch analysis across multiple plates.

Its experiment wizard contains ready-to-use acquisition and analysis

templates for key applications, and lets you rapidly define and store

your own templates. One-click instrument maintenance protocols

make it easy to keep the bioanalyzer performing optimally.

BD Biosciences assays for the BD FACSArray bioanalyzer are devel-

oped with consideration for simplicity and small sample volume,

while taking advantage of the analyzer’s multiparameter detection

capability which results in many answers per sample.

System speed, novel software tools, and streamlined assay formats

accelerate your total productivity and interpretation of high-content

biological data for the answers you need.

Discover the Flexibility

The BD FACSArray bioanalyzer has an optical detection system that

is ideal for multicolor flow cytometry experiments. The optics are

capable of collecting up to four fluorescent colors from each sample,

as well as forward and side scatter light. Correlated forward and

side scatter signals are crucial for differentiating events of interest

from interfering debris. With its multiparameter capabilities, you can

resolve various populations within a single cellular or bead sample.

For multiplex bead applications, antibody-labeled fluorescent beads

capture and quantify secreted proteins or proteins in cell lysates

using BD CBA technology. With BD CBA Flex Set assays, you tailor

the bead array to your specific experiment. For cellular applications,

antibodies conjugated to fluorophores bind specifically to cell surface

or intracellular proteins. This technique can be applied to resting,

immune-challenged, or drug-exposed samples to differentiate cells

and to define their biological responses. Additionally, dyes can be

used to determine cell viability. BD Biosciences applications cover the

entire range of species including studies of human, murine, dog, and

non-human primate samples.



Imaging and High Content Analysis (continued)

*ClassI(1)laserproduct†Forinvitrodiagnosticuse‡Seven-andeight-colorimmunophenotypingrequirevalidationbyuser.§Forresearchuseonly.Notforuseindiagnosticortherapeuticprocedures.

The BD™ High Throughput Sampler (HTS) option available for the BD FACSCanto™ II,§ BD™ LSR II, and BD FACSCalibur™ flow cytometers

provides fully automated and rapid sample acquisition from either 96- or 384-well microtiter plates.

• Under 15-minute acquisition for a 96-well tray

• Carryover <1% in high throughput mode, <0.5% carryover in standard mode

• User-adjustable mixing, wash, and acquisition parameters

• Adaptable to a wide range of applications

• Robotic-accessible design

BD Flow Cytometry Instrumentation

BD FACSArray bioanalyzer* Inquire

•Fastmicrotiterplatesampler

•Six-parameterdetection(twoscatterandfourfluorescences)

•Intuitivesoftware

•Digitalsignalprocessingwithupto15,000eventspersecond

•Compact,affordablebenchtopunit

BD FACSCalibur flow cytometer* Inquire

•Theindustrystandardindual-laser,six-parameter,four-colorflowcytometricanalysis

•IncludesBDCellQuest™ProSoftware

•BDFACSCaliburandBDCellQuestProareforInVitroDiagnosticUsewhenusedwithIVDclearedassays

•BDHTSoption

BD FACSCanto II flow cytometer*† Inquire

•Three-laserinstrumentwithtrueeight-colorcapability‡

•Powerfuldigitalsoftwaretomakemulticolorexperimentationfastandsimple

•Resolvesthedimmesteventsbyincreasedsensitivity

•BDHTSoption‡§

BD LSR II flow cytometer* Inquire

•Incorporatesdigitalelectronicsforusewithuptofourfixed-alignmentlasers(488nm,638nm,405nm,andUV)

•Detects18colorsthrougharevolutionarynewopticaldesign

•Flexibleandmodularforfutureupgrades

•BDHTSoption

BD FACSAria cell sorting system* Inquire

•Firstbenchtophigh-speedsorterwithfixed-alignmentcuvetteflowcell

•Cuvetteflowcellforsuperiorfluorescencesensitivity

•Uptothreeair-cooledlasersat488,633,and405nmwavelengths

•Digitalacquisitionratesofupto70,000eventspersecond

•Multicoloranalysisofupto15parameters

•Two-andfour-waybulksortingdevicesforavarietyoftubesizes

•OptionalBD™AutomatedCellDepositionUnit(ACDU)forsortingtoBD™Multiwellplatesormicroscopeslides

instrument features cat.no.



1. Histone deacetylase inhibitors arrest polyglutamine-dependent neurodegeneration in Drosophila. Nature 18;413(6857):739-43 (2001).

2. Chilkoti, A., et al., Investigating the relationship between surface chemistry and endothelial cell growth: partial least-squares regression of the static secondary ion mass spectra of oxygen containing plasma-deposited films. Analytical Chemistry 67:2883 (1995).

3. Sotiropoulou, P, et al., Characterization of the Optimal Culture Condition for Clinical Scale Production of Human Mesenchymal Stem Cells. Stem Cells 24: 462-471 (2006).

Additional References

Chat, G. and M. Coca-Prados. Comparison of tissue culture systems for the growth of ciliary epithelial cells. BDL Monograph (1986).

Chinn, J.A., et al., Laboratory preparation of plasticware to support cell culture. J. Tissue Culture Methods 16:155 (1994).

4. Ertel, S., et al., Endothelial cell growth on oxygen-containing films deposited by radiofrequency plasmas; the role of surface carbonyl groups. Biomater. Sci.: Polym. Ed. 3:163 (1991).

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