Cell Signaling Technology

Product Pathways - DNA Damage

53BP1 Antibody #4937

Applications Reactivity MW (kDa) Source
W IP IHC-P IF-IC F H Mk 450 Rabbit

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

Specificity / Sensitivity

53BP1 Antibody detects endogenous levels of total 53BP1 protein independent of phosphorylation.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to residues near the center of human 53BP1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HT29 cells, using 53BP1 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using 53BP1 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma, using 53BP1 Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using 53BP1 Antibody in the presence of control peptide (left) or antigen-specific peptide (right).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HT29 cells, using 53BP1 Antibody (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using 53BP1 Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).


Background

p53-binding protein 1 (53BP1) was originally identified as a p53 binding partner that could enhance the transcriptional activity of p53 (1,2). 53BP1 consists of two BRCA1 carboxy terminal (BRCT) domains that allow for binding to p53 and a separate domain responsible for binding to phosphorylated histone H2A.X (3). 53BP1 rapidly translocates to nuclear foci following treatment of cells with ionizing radiation (IR) or radiomimetic agents that cause DNA double strand breaks (DSBs) (4,5). Because of this localization to DSBs and homology to the yeast protein Rad9, a role for 53BP1 in DSB repair has been proposed. Recruitment of 53BP1 to sites of DNA damage has been demonstrated to be independent of ATM, NBS1, and DNA-PK (4) and retention of 53BP1 at DNA breaks requires phosphorylated H2A.X (6). In cells lacking 53BP1, phosphorylation of ATM substrates is reduced, suggesting that 53BP1 is upstream of ATM (7). In response to IR, phosphorylation of 53BP1 at serines 6, 25, 29, and 784 by ATM has been demonstrated, but phosphorylation at these sites is not required for localization of 53BP1 to sites of DSBs (6).

  1. Iwabuchi, K. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 6098-6102.
  2. Iwabuchi, K. et al. (1998) J. Biol. Chem. 273, 26061-26068.
  3. Mochan, T.A. et al. (2004) DNA Repair (Amst) 3, 945-952.
  4. Schultz, L.B. et al. (2000) J. Cell Biol. 151, 1381-1390.
  5. Anderson, L. et al. (2001) Mol. Cell. Biol. 21, 1719-1729.
  6. Ward, I.M. et al. (2003) J. Biol. Chem. 278, 19579-19582.
  7. DiTullio, R.A. et al. (2002) Nat. Cell Biol. 4, 998-1002.

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