Product Pathways - Chromatin Regulation

Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb #9718

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Applications	Reactivity	Sensitivity	MW (kDa)	Isotype
W IHC-P IF-IC F	H M R Mk	Endogenous	15	Rabbit IgG

Applications Key: W=Western Blotting IHC-P=Immunohistochemistry (Paraffin) IF-IC=Immunofluorescence (Immunocytochemistry) F=Flow Cytometry
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey
Species cross-reactivity is determined by Western blot.

Protocols

9718:: Flow, IHC / Paraffin, Western Blotting

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.

Western Blotting

Western blot analysis of extracts from untreated or UV-treated 293 cells, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (upper) or Histone H2A.X Antibody #2595 (lower).

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HT-29 cells untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb in the presence of control peptide (left) or Phospho-Histone H2A.X (Ser139) Blocking Peptide #1260 (right).

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb, showing nuclear localization.

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma untreated (left) or lambda-phosphatase-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb.

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) or UV-treated (green), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb compared to a nonspecific negative control antibody (red).

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or UV-treated (right), using Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (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. (1998) Annu. Rev. Biochem. 67, 545-579.
Hansen, J. C. et al. (1998) Biochemistry 37, 17637-17641.
Cheung, P. et al. (2000) Cell 103, 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

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.