F eat u r es & B e n eF i ts

Simple protocolS

multiple interchangeable FluorophoreS

long-term live cell imaging

convenient multiplexing

more inFormation

High-Content Screeningwith the HaloTag® platform

✓ One HaloTag® protein construct can be imaged with interchangeable fluorophores

✓ Conduct long-term screens in live cells as the ligand is covalently bound to HaloTag

✓ Easily multiplex with antibody- or GFP-based imaging

H i g H - C o n t e n t s C r e e n i n g w i t H t H e H a l o ta g ® p l at F o r m

FeatureS & beneFitS

s i m p l e p roto Co l s

multiple interchangeable FluorophoreS

long-term live cell imaging

convenient multiplexing

more inFormation

Simple cell labeling protocols

HaloTag

HaloTag

1. Transfect HaloTag® construct

3. Image

Express protein

OR

2a. Add HaloTag TMR- or R110Direct™ ligand (no wash)

2b. Add HaloTag ligand (15 min. label, 30 min. wash)

9758

MA

H i g H - C o n t e n t s C r e e n i n g w i t H t H e H a l o ta g ® p l at F o r m

FeatureS & beneFitS

Simple protocolS

m u lt i p l e i n t e rC H a n g ea B l e F lu o ro p H o r es

long-term live cell imaging

convenient multiplexing

more inFormation

Multiple interchangeable fluorophoresHaloTag® ligands are bright, stable and specific

Confocal imaging of labeled Halotag® fusion proteins in cells with tmrDirect™ and r110Direct™ ligands.

labeling with Halotag fluorescent ligands is highly specific to Halotag, and stable through fixation and freeze-thaw.

H i g H - C o n t e n t s C r e e n i n g w i t H t H e H a l o ta g ® p l at F o r m

FeatureS & beneFitS

Simple protocolS

multiple interchangeable FluorophoreS

lo n g-t e r m l i v e C e l l i m ag i n g

convenient multiplexing

more inFormation

Long-term live cell imaging

48 hours of cell division with incorporated Halotag® ligand

confocal microscopy of u2oS cells expressing halotag-nlS. plate and label cells with a single addition of ligand prior to imaging. robust cell division and lack of cell toxicity after 48 hours.

9766

CA

H i g H - C o n t e n t s C r e e n i n g w i t H t H e H a l o ta g ® p l at F o r m

FeatureS & beneFitS

Simple protocolS

multiple interchangeable FluorophoreS

long-term live cell imaging

Co n v e n i e n t m u lt i p l e x i n g

more inFormation

Multiplex with other labeling methods

Halotag ligands can…• multiplex with antibody staining and/or gFp• remain bright and stable after cell fixation• undergo freeze/thaw

5379

CA

4990

CC

halotag® with gFp halotag with antibodies

• halotag–nlS labeled with tmr ligand

• expression of a gFp-a-tubulin

• p65-halotag labeled with tmr ligand

• Fixed cells stained with anti-a-tubulin primary ab, and alexa-488-conjugated secondary ab

live-cell imaging Fixed-cell imaging

H i g H - C o n t e n t s C r e e n i n g w i t H t H e H a l o ta g ® p l at F o r m

FeatureS & beneFitS

Simple protocolS

multiple interchangeable FluorophoreS

long-term live cell imaging

convenient multiplexing

m o r e i n Fo r m at i o n

promega corporation • 2800 WoodS holloW road • madiSon, Wi 53711-5399 uSa • telephone 608-274-4330 www.promega.com • ©2011 promega corporation • all rightS reServed • priceS and SpeciFicationS Subject to change Without prior notice • printed in uSa 2/11 • 19771 • part #iS038

References

huybrechts, S.j. et al. (2009) peroxisome dynamics in cultured mammalian cells. Traffic 10(11): 1722-33.

Svendsen, S. et al. (2008) Spatial separation and bidirectional trafficking of proteins using a multi-functional reporter. BMC Cell Biol. 9: 17.

Yamaguchi, K. et al. (2009) pulse-chase experiment for the analysis of protein stability in cultured mammalian cells by covalent fluorescent labeling of fusion proteins. Methods Mol. Biol. 577: 121-31.

los, g.v. et al. (2008) halotag: a novel protein labeling technology for cell imaging and protein analysis. ACS Chem. Biol., 3(6): 373-82.

Ordering Informationp r o D u C t s i z e C ata l o g #

Halotag® F luorescent l igands various g8251, g8252…

Halotag® react ive l igands 5mg p6751, p1691…

anti -Halotag® pab 200µg g9281

halotag is a registered trademark of promega corporation. halolink, tmrdirect, and r110direct are trademarks of promega corporation.