p53 AntibodyProduct information
Product Pathways - Cell Cycle / Checkpoint
p53 Antibody #9282
|9282S||100 µl (10 western blots)||---||In Stock||---|
|9282T||20 µl (2 western blots)||---||In Stock||---|
|9282||carrier free and custom formulation / quantity||email request|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP
Directions For Use
For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Specificity / Sensitivity
p53 Antibody recognizes endogenous levels of total p53 protein. This antibody binding has been mapped to the amino terminus and DNA binding domain of human p53 protein. This antibody does not cross-react with with other p53-related proteins.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a full-length human p53 fusion protein. Antibodies are purified by protein A and peptide affinity chromatography.
Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells treated with UV (100 J/m2 followed by a 3 hour recovery) and either p53 Antibody or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CDKN1A Promoter Primers #6449, human MDM2 intron 2 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis (1). p53 is phosphorylated at multiple sites in vivo and by several different protein kinases in vitro (2,3). DNA damage induces phosphorylation of p53 at Ser15 and Ser20 and leads to a reduced interaction between p53 and its negative regulator, the oncoprotein MDM2 (4). MDM2 inhibits p53 accumulation by targeting it for ubiquitination and proteasomal degradation (5,6). p53 can be phosphorylated by ATM, ATR, and DNA-PK at Ser15 and Ser37. Phosphorylation impairs the ability of MDM2 to bind p53, promoting both the accumulation and activation of p53 in response to DNA damage (4,7). Chk2 and Chk1 can phosphorylate p53 at Ser20, enhancing its tetramerization, stability, and activity (8,9). p53 is phosphorylated at Ser392 in vivo (10,11) and by CAK in vitro (11). Phosphorylation of p53 at Ser392 is increased in human tumors (12) and has been reported to influence the growth suppressor function, DNA binding, and transcriptional activation of p53 (10,13,14). p53 is phosphorylated at Ser6 and Ser9 by CK1δ and CK1ε both in vitro and in vivo (13,15). Phosphorylation of p53 at Ser46 regulates the ability of p53 to induce apoptosis (16). Acetylation of p53 is mediated by p300 and CBP acetyltransferases. Inhibition of deacetylation suppressing MDM2 from recruiting HDAC1 complex by p19 (ARF) stabilizes p53. Acetylation appears to play a positive role in the accumulation of p53 protein in stress response (17). Following DNA damage, human p53 becomes acetylated at Lys382 (Lys379 in mouse) in vivo to enhance p53-DNA binding (18). Deacetylation of p53 occurs through interaction with the SIRT1 protein, a deacetylase that may be involved in cellular aging and the DNA damage response (19).
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- Shieh, S.Y. et al. (1997) Cell 91, 325-34.
- Chehab, N.H. et al. (1999) Proc Natl Acad Sci U S A 96, 13777-82.
- Honda, R. et al. (1997) FEBS Lett 420, 25-7.
- Tibbetts, R.S. et al. (1999) Genes Dev 13, 152-7.
- Shieh, S.Y. et al. (1999) EMBO J 18, 1815-23.
- Hirao, A. et al. (2000) Science 287, 1824-7.
- Hao, M. et al. (1996) J Biol Chem 271, 29380-5.
- Lu, H. et al. (1997) Mol Cell Biol 17, 5923-34.
- Ullrich, S.J. et al. (1993) Proc Natl Acad Sci U S A 90, 5954-8.
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- Lohrum, M. and Scheidtmann, K.H. (1996) Oncogene 13, 2527-39.
- Knippschild, U. et al. (1997) Oncogene 15, 1727-36.
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- Ito, A. et al. (2001) EMBO J 20, 1331-40.
- Sakaguchi, K. et al. (1998) Genes Dev 12, 2831-41.
- Solomon, J.M. et al. (2006) Mol Cell Biol 26, 28-38.
- Russell, J. L. et al. (2002) Mol. Cell. Biol. 22, 1360-1368. Applications: Western Blotting.
- Stansel, R. M. et al. (2002) J. Biol. Chem. 277, 11625-11628. Applications: Immunological Electron Microscopy.
- Vaghefi, H. and Neet, K.E. (2004) Oncogene 23, 8078-8087. Applications: Western Blotting.
- Daniely, Y. et al. (2002) Mol. Cell. Biol. 22, 6014-6022. Applications: IP, Western Blotting.
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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.
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