Chromatin Regulation / Acetylation
Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (Alexa Fluor® 647 Conjugate) #9720
| Item # | Description | List Price | Web Price | QTY | Buy |
|---|
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| IF-IC F | H M | Endogenous | Rabbit |
Applications Key:
IF-IC = Immunofluorescence (Immunocytochemistry) F = Flow Cytometry
Reactivity Key:
H = Human M = Mouse
Species cross-reactivity is determined by Western blot.
Protocols
- 9720
- Flow, Immunofluorescence,
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 antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser139 of human H2A.X. The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-5. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.
Description
This Cell Signaling Technology Antibody was conjugated to Alexa Fluor®647 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®647 conjugate) will also recognize Phospho-Histone H2A.X in these species.
Flow Cytometry
Flow cytometric analysis of Jurkat cells, untreated (green) or etoposide-treated (blue), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (Alexa Fluor® 647 Conjugate).
IF-IC
Confocal immunofluorescent analysis of NIH/3T3 cells, untreated (left) or etoposide-treated (right), double-labeled with Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue) and beta-Tubulin Antibody #2146 (red).
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).
- Workman, J.L. and Kingston, R.E. et al. (1998) Annu. Rev. Biochem.67, 545 - 579.
- Hansen, J. C. et al. (1998) Biochemistry37, 17637 - 17641.
- Cheung, P. et al. (2000) Cell103, 263 - 271.
- Thorne, A. W. et al. (1990) Eur. J. Biochem.193, 701 - 713.
- Rogakou, E. P. et al. (1998) J. Biol. Chem.273, 5858 - 5868.
- Burma, S. et al. (2001) J. Biol. Chem.276, 42462 - 42467.
- Rogakou, E. P. et al. (1999) J. Cell Biol.146, 905 - 916.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 2577 Phospho-Histone H2A.X (Ser139) Antibody
- 9708 Phospho-Histone H3 (Ser10) Antibody (Alexa Fluor® 488 Conjugate)
- 2595 Histone H2A.X Antibody
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.
