nordic-mubio vet flyer update april 2015valterocchiena.com/pdf/nordic-mubio.pdf · antibodies for...
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Nordic MUbio
Visit www.nordicmubio.com/Products now to �nd the
products you need for your immunological research, or
get in touch on [email protected]
FIX&PERM® is a simple procedure making use of
two reagents. Reagent A gently �xes cells, while
Reagent B permeabilizes them.
The speci�c formulation of FIX&PERM reduces
background while allowing the simultaneous addition of
permeabilization medium and �uorochrome labelled
antibodies for staining of membrane proteins as well as
intracellular components, such as cytoplasmic enzymes,
nuclear proteins, oncoproteins, cytokines,
immunoglobulins, and so on.
• Mildly ixes cells, preserving their low cytometric scatter
characteristics
• Allows simultaneous characterisation of both
intracellular and cell surface markers
• Rapid technique – whole procedure can be carried out in
less than one hour, ready for immediate analysis or storage
for 24 hours
• Stringent QC procedures – the quality of each lot is
determined using well-de�ned blood samples and
subsequent comparison of immunostaining and scatter
characteristics of obtained leukocyte populations, ensuring
consistent and reliable results lot after lot
• A range of antibodies to intracellular and membrane
antigens with optimised protocols for use with FIX&PERM®
FIX&PERM®Cell Fixation and
Permeabilization Kit
Reagent A gently �xes cells preserving
their forward and side-scatter
characteristics
Reagent B permeabilizes the cell
Allowing binding and detection of
intracellular antigens
Reagent B and labelled antibodies are
then added
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here:
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The majority of available monoclonal antibody conjugates can be used with
FIX&PERM®.
However, in some cases the �xation step may require optimization.
Nordic-MUbio o�ers a number of mouse anti-human conjugated (PE/FITC)
monoclonal antibodies pre-optimised for use with FIX&PERM®.
FIX&PERM®Cell Fixation
and Permeabilization Kit
More products with protocols optimised for FIX&PERM can be found here:
http://www.nordicmubio.com/Services/FIXPERM
All FIX&PERM® kits are CE Marked
Procedure
For each sample to be analysed use
50 µl of whole blood, bone marrow
or mononuclear cell suspension in a
5 ml tube
Size Cat. Number
Fix&PERM® Kit 2 x 20 mL GAS-002
Fix&PERM® Sample Kit 2 x 5 mL GAS-002M
Fix&PERM® 1000 Kit 2 x 100 mL GAS-002-1
Fix&PERM® Reagent A (Fix) 1 x 100 mL GAS-002A-1
Fix&PERM® Reagent B (Perm) 1 x 100 mL GAS-002B-1
Add 100 µl of Reagent A (Fixation
Medium, stored and used at room
temperature)
Add 5 ml phosphate bu�ered saline
and centrifuge cells for 5 minutes at
300 g
Remove supernatant and add to cell
pellet 100 µl Reagent B and 20 µl of
the appropriate monoclonal antibody
Wash cells with PBS as above
Remove supernatant and resuspend
cells in sheath luid for immediate
analysis
Alternativly, resuspend cells in 0.5 ml
1.0 % formaldehyde and store at
2-8°C in the dark.
Analyse cells within 24 hours
Vortex at low speed for 1-2 seconds
Incubate for 15 minutes at room
temperature
Incubate for 15 minutes at room
temperature
Antigen Isotype Size Cat. Number
CD3 IgG1 0.2 mg GM-4011
100 tests GM-4012 (FITC)
100 tests GM-4013 (PE)
CD22 IgG1 0.2 mg GM-4051
100 tests GM-4052 (FITC)
100 tests GM-4053 (PE)
Mouse anti-Human Combi Products
MPO-C2 IgG1 0.2 mg GM-4191
100 tests GM-4192 (FITC)
100 tests GM-4193 (PE)
IgG Negative Control (FITC) and IgG Negative Control (PE) GIC-201
Anti-Lysozyme (FITC) and anti-Lactoferrin (PE) GIC-206
Anti-Myeloperoxidase-C2 (FITC) and anti-Lactoferrin (PE) GIC-212
Anti-Myeloperoxidase-C2 (FITC) and anti-CD22 (PE) GIC-214
Anti-Myeloperoxidase-C2 (FITC) and anti-CD3 (PE) GIC-213
Catalogue
NumberDescription Format
Unit
Size
Bii-nGFPab2-100Rabbit anti-GFP
(native)
Ammonium Sulfate
fraction0.1 mg
Bii-nGFPab3-100Rabbit anti-GFP
(native)Protein A Puriied 0.1 mg
Bii-nGFPab4-100Rabbit anti-GFP
(native)Ainity Puriied 0.1 mg
Bii-nGFPab4-PORabbit anti-GFP
(native):PO
Soyabean Peroxidase
Conjugate0.1 ml
Bii-nGFPabC1-100Chicken anti-GFP
(native)
Ammonium Sulfate
fraction0.1 mg
Bii-rGFP-100
Recombinant
Green Fluorescent
Protein (wild type)
Recombinant Protein 0.1 mg
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Green Fluorescent Protein
(GFP) is a 27kD protein derived
from the Aequorea victoria (A.
victoria) jellyish. It is a naturally
luorescent protein which has
revolutionised life science and
has been used to study proteins
in a wide range of applications.
GFP has been used extensively
to monitor gene expression; to
localise proteins at the cellular
and sub-cellular level; to track
metastatic cells and as a tag to
identify proteins in vitro.
The enormous interest in GFP
as a protein marker has given
rise to the development of
polyclonal and monoclonal
antibodies against GFP, used
for the immunolocalisation
and immunoblotting of GFP-
fused proteins in a variety of
applications.
Since its discovery, many variants
of GFP have been engineered
making antibodies recognising a
wide range of these variants an
important, and useful, scientiic
tool for use alongside GFP-fused
proteins.
A. victoria is a bioluminescent jellyish, or
hydromedusa, found of the west coast
of North America.
Nordic-MUbio is pleased to announce the release
of a new range of products directed against the
native form of GFP.
Our new range of anti-GFP antibodies bring a number of advantages
over antibodies produced against recombinant, or variant, forms of
GFP.
This includes the ability to detect GFP produced from a wide range
of diferent sources, including native, recombinant, and a number of
diferent mutants, without sufering from high levels of non-speciic
binding/background.
Part of the reason many anti-recombinant GFP antibodies sufer from
high background issues is due to contamination of an innoculum
with other E.coli components, leading to increased non-speciic
binding.
Our products were tested for cross-reactivity to a wide range of
antigens, including many derived from E.coli, and demonstrated no
cross-reactivity.
High Quality Green Fluorescent
Protein (GFP) Antibodies
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Anti-GFP
Ab’ source
HPLC
Purity
SDS
Purity
Western
Speciicity
Native GFP
AntibodyClean Clean Clean
Competitor A Contaminants ContaminantsSome
Background
Competitor B Clean CleanHigh
Background
Competitor C Multiple peaksSlight
Contaminantsn/a
Competitor D Many peaks
(contains BSA)Not run n/a
Competitor E Multiple peaks Contaminants n/a
Competitor F Multiple peaks Contaminants n/a
Competitor G Multiple peaks Clean n/a
Example Size Exclusion
Chromatography (HPLC) Results
Competitor D
Bii-nGFPab3-100
(Protein A puriied)
Competitor E
Western blot Anlaysis
The gels in these images have been loaded
with E.coli control, not expressing GFP
(lane A), E.coli expressing GFP (lane B), and
molecular weight standards (lane C).
Competitor A Competitor B Nordic-MUbio
Native GFP
antibody
These images show intact GFP (main band
at 27 kDa), with some high and low mol.
weight bands related to GFP present in
all. The antisera from competitors A and B
show more background reactivity in the E.
coli control (lane A).
To ensure Nordic-MUbio’s new GFP antibody range is of the highest
quality a number of comparison experiments were run with the best-
selling antibodies from other suppliers against recombinant GFP,
assessing purity, speciicity, and titre of the antibodies.
This page presents a number of the indings from these anlayses.
Visit www.nordicmubio.com/Products now to ind the
products you need for your immunological research, or
get in touch on [email protected]
The HPLC analyses of the Nordic-
MUbio ainity puriied antibodies
show just a single peak, illustrating
their purity. The competitor
antibodies with multiple peaks
have contaminants, such as serum
albumin, which can afect the results
of e.g. Western blots or ELISAs.
Bii-nGFPab3-100
(Protein A puriied)
How do we know our GFP products are
the highest quality?
Isotyping in research
In vaccine research, the isotype of the antibodies produced post-vaccination can give information on the type of immune response induced. For example, stronger induction of an IgA response might be desirable for a vaccine targeting viral pathogens, induc-ing mucosal immunity. While higher levels of IgG2 are corre-lated with a Th1 (pro-in�ammatory) response, and higher IgG1 is more characteristic of a Th2 (anti-in�ammatory) response.
Screening for antibodies devel-oped against biologic drugs is a requirement of most clinical trials and post-approval monitoring. When a patient does develop an immune response it can be useful to determine both the speci�city and antibody isotype of the response.
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Nordic MUbio
Nordic-MUbio o�ers the highest quality
range of products for the study and
analysis of human immunoglobulins.
Our highly validated product range spans mouse monoclonal
antibodies against all isotypes of human immunoglobulins,
including allotypes, and are available in multiple conjugates
suitable for applications such as ELISA, �ow cytometry, western
blot and immunohistochemistry.
We also have a range of highly puri�ed human immunoglobulins,
suitable for use as references and antigens.
Determining the class (isotype) and subclass of an antibody is
important for a variety of reasons, from selection of puri�cation
protocols to day-to-day immunological methods and clinical
diagnostics.
Di�erent antibody classes and subclasses serve a variety of di�erent
functions within the body and can be indicative of disease states
and immunological processes. For example, IgE is characteristic of
allergic reactions. Each antibody isotype has unique e�ector
functions, capable of binding di�erent cellular receptors.
Visit nordicmubio.com/Products now to �nd the answer to all
your human immunoglobulin needs, or get in touch on
Human Immunoglobulins
Contact your local distributor
here:
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Nordic MUbio
Nordic-MUbio has developed an
ever-expanding range of high
quality immunological research
tools, manufactured to strict ISO
9001 guidelines, to provide both
reassurance and the best possible
tools for your research.
Visit nordicmubio.com/Products
now to �nd the answer to all your
human immunoglobulin needs,
or contact us:
Puri�ed Human
ImmunoglobulinCat. Number
IgG HuIgG
IgG subclasses standard Serum NOR-01
IgG1 lambda G1m(f) P 106
IgG4 kappa Gm4a P 182
IgG1 lambda G1m (a) P 210
IgG3 lambda G3m (g) P 289
IgG4 kappa Gm4b P 304
IgG2 lambda G2m n-) P 315
IgG3 kappa G3m(b) P 411
IgG2 lambda G2m (n+) P 532
IgA subclasses standard Serum NOR-04
IgA1 lambda (dimer) P 444
Secretory IgA P 020
IgA1 kappa, polymer P 430
IgA2 lambda A2m (1-, 2+) P 505
IgA2 (1+,2-) lambda P 237
Human IgE kappa P 050
Human Bence Jones kappa
protein (Free light chain, kappa type)
P 016
Human Bence Jones lambda
protein (Free light chain, lambda type)
P017
Human Immunoglobulin Products
Mouse anti-human Cat. Number
IgG1 MAHu/IgG1 B,H,F,T
IgG2 (Fab subclass speci�c) MAHu/IgG2 B,H,F,T
IgG2 (Fc subclass speci�c) MAHu/IgG2Fc B,H,F,T
MAHu/IgG3 B,H,F,T
MAHu/IgG3m(u) B,H,F,T
IgG4 (pFc subclass speci�c) MAHu/IgG4 B,H,F,T
IgG (class speci�c) MAHu/IgGc B,H,F,T
IgM (class speci�c) MAHu/IgM B,H,F,T
IgA1 MAHu/IgA1 B,H,F,T
IgA2 MAHu/IgA2 B,H,F,T
IgA2 (m2 allotype) MAHu/IgA2(m)2 B,H,F,T
IgA1 + IgA2 MAHu/IgAc B,H,F,T
IgE MAHu/IgE B,H,F,T
IgD MAHu/IgD B,H,F,T
*B= Biotin, H=Horseradish peroxidase, F = FITC, T=TRITC
AvailableConjugates*
IgG3 (F(ab'')2 subclass speci�c)
IgG3 (G3m(U) allotype speci�c)
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Nordic MUbio
Visit www.nordicmubio.com/Products now to �nd the
products you need for your immunological research, or
get in touch on [email protected]
Nordic-MUbio has developed an
ever-expanding range of high quality
immunological research tools,
manufactured to strict ISO 9001
guidelines, to provide both
reassurance and the best possible
tools for your research.
Veterinary Immunoglobulins
Nordic-MUbio is dedicated to helping veterinary
scientists advance their understanding of animal
immunology, helping to safeguard everything
from domestic pets to livestock in health and
disease.
Our highly validated veterinary immunoglobulin products
include a wide range of both host and target species, giving
researchers the scope and choice they need.
As in humans, animal immunoglobulins play a huge role in
health and disease, with di�erent antibody classes and
subclasses indicative of di�erent disease states or normal
activity of immunological processes.
This makes animal immunoglobulins of interest and
importance to a huge range of research areas from infectious
diseases to cancer research.
We know your research needs reliable products that just work.
And that’s why we’ve developed a portfolio of veterinary
immunoglobulin products with a wealth of validation data,
suitable for use as either primary or secondary antibodies and
in a range of formats to suit your research needs.
Whether your application is immunoprecipitation, ELISA or
immunohistochemistry, if we list it as an application you can
be sure that our products will work. Saving you not only
money, but more importantly, time and precious sample.
Together with our strict ISO 9001 manufacturing and quality
control you can be sure each of our products will work time
after time, lot after lot.
Contact your local distributor
here:
Our 15 Top Selling Veterinary Immunoglobulin Products
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Conjugate Key:B: Biotin
H: Horseradish peroxidase
F: FITC
T: TRITC
Applications Key:EL: ELISA
ICC: Immunocytochemistry
IHC(F): Immunohistochemistry (frozen)
IHC(P): Immunohistochemistry (parra�n)
DB: Dot blotting
IB: Immunoblotting
IP: Immunoprecipitation
IF: (in)direct immuno�uorescence
Our complete range of veterinary
products includes reagents for research
into: Cow, Dog, Chicken, Duck, Electric
Eel, Cat, Goat, Guinea Pig, Hamster,
Horse, Mouse, Rabbit, Rat, Sheep, Snake,
Swine and many more.
Anti IgG*
Host Anti- Cat. Number Conjugate Application
Anti IgG1, IgG2a, IgG2b, IgG3, IgA, IgM*
Host Anti- Cat. Number Conjugate Application
Anti IgG1, IgG2a, IgG2b, IgG3*
Host Anti- Cat. Number Conjugate Application
Anti serum proteins
Host Anti- Cat. Number Conjugate Application
Rabbit Goat RAG/IgG(H+L) B, F, H, T EL, ICC, IHC(P), DB, IB, IP
Swine Mouse SwAM/IgG(H+L) F EL, ICC, IHC(F), IF
Rabbit Swine RASw/IgG(H+L) B, F, H, T EL, ICC, IHC(P), DB, IB, IP, IF
Rabbit Dog RAD/IgG(H+L) B, F, H, T EL, ICC, IHC(P), DB, IB, IP
Goat Cow GAB/IgG(H+L) B, F, H EL, ICC, IHC(P), IHC(F), DB, IB, IP
Goat Rabbit GAR/IgG(H+L) B, F, H, T
B, F, H, T
EL, ICC, IHC(P), IHC(F), DB, IB, IP
Rabbit Horse RAHo/IgG(H+L) B, F, H EL, ICC, IHC(P), IHC(F), DB, IB, IP
Rabbit Cat RACa/IgG(H+L) B, F, H EL, ICC, IHC(F), DB, IB, IP, IF
Rabbit Duck RADu/IgG(H+L) B, F, H EL, ICC, IHC(P), IHC(F), DB, IB, IP
Rabbit Chicken RACh/IgG(H+L) B, F, H EL, ICC, IHC(P), DB, IB, IP, IF
Anti IgD (Fc speci�c)
Host Anti- Cat. Number Conjugate Application
Goat Mouse GAM/IgD(Fc) EL, ICC, IHC(F), IP, IF
B, F, H, TRabbit Mouse RAM/Ig EL, ICC, IHC(P), IHC(F), DB, IB, IP
B, F, H, TGoat Mouse GAM/IgG(H+L) EL, ICC, IHC(P), DB, IB, IP
_Rabbit Horse RAHo/TSP IP
Anti Beta lactoglobulin
Host Anti- Cat. Number Conjugate Application
_Rabbit Cow RAB/BLgl IP
*Heavy and Light chains