Product Pathways - Chromatin Regulation

Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (Alexa Fluor® 488 Conjugate) #9719

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Applications	Reactivity	Sensitivity	Isotype
IF-IC F	H M	Endogenous	Rabbit IgG

Applications Key:  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb detects endogenous levels of H2A.X only when phosphorylated at serine 139.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser139 of human H2A.X. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-5.

Description

This Cell Signaling Technology Antibody was conjugated to Alexa Fluor®488 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody #9718 reacts with human and mouse Phospho-Histone H2A.X. CST expects that Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (Alexa Fluor® 488 conjugate) will also recognize Phospho-Histone H2A.X in these species.

Background

Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation and methylation (2-4). These modifications occur in response to cell signaling stimuli and have a direct effect on gene expression. DNA damage caused by ionizing radiation, UV-light, or radiomimetic agents results in rapid phosphorylation of the histone H2A family member H2A.X at Ser139 by ATM (5,6). Within minutes following DNA damage, Ser139-phosphorylated H2A.X localizes to sites of DNA damage at subnuclear foci (7).

1. Workman, J.L. and Kingston, R.E. (1998) Annu. Rev. Biochem. 67, 545-579.
2. Hansen, J. C. et al. (1998) Biochemistry 37, 17637-17641.
3. Cheung, P. et al. (2000) Cell 103, 263-271.
4. Thorne, A. W. et al. (1990) Eur. J. Biochem. 193, 701-713.
5. Rogakou, E. P. et al. (1998) J. Biol. Chem. 273, 5858-5868.
6. Burma, S. et al. (2001) J. Biol. Chem. 276, 42462-42467.
7. Rogakou, E. P. et al. (1999) J. Cell Biol. 146, 905-916.

Application References

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Companion Products

• 2577 Phospho-Histone H2A.X (Ser139) Antibody
• 9716 Phospho-Histone H3 (Ser10) Antibody (Alexa Fluor® 647 Conjugate)

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.