Cell Signaling Technology

Product Pathways - Chromatin Regulation

Phospho-Histone H2A.X (Ser139) Antibody #2577

Applications Reactivity MW (kDa) Source
W IHC-P IF-IC F H M R 15 Rabbit

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-H2A.X (Ser139) Antibody detects endogenous levels of H2A.X only when phosphorylated at Ser139.

Source / Purification

Antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser139 of human H2A.X.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or UV-treated, using Phospho-Histone H2A.X (Ser139) Antibody (upper) or Histone H2A Antibody #2572 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HT-29 cells untretaed (left) or treated (right) with UV (upper) or doxorubicin (lower) using Phospho-Histone H2A.X (Ser139) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded L36 pancreatic adenocarcinoma xenografts, untreated (left) or chemotherapy treated (right). (High magnification inset) (Tissue provided by Dr. Murray Resnick, Rhode Island Hospital).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast tumor control (left) or lambda phosphatase-treated (right) using Phospho-Histone H2A.X (Ser139) Antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) and UV-treated (green), using Phospho-Histone H2A.X (Ser139) Antibody compared with a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal microscopic images of HeLa cells, UV treated (A) and untreated (B), showing nuclear stain with Phospho-Histone H2A.X (Ser139) Antibody (red) and Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb #9255 (green).


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.

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