Product Pathways - DNA Damage
Phospho-p53 (Ser15) Antibody #9284
|9284L||300 µl (30 western blots)||---||In Stock||---|
|9284S||100 µl (10 western blots)||---||In Stock||---|
|9284P||40 µl (4 western blots)||---||In Stock||---|
|9284||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat, Monkey||Endogenous||53||Rabbit|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Mink, Bovine, Pig.
Specificity / Sensitivity
Phospho-p53 (Ser15) Antibody detects endogenous levels of p53 only when phosphorylated at serine 15. The antibody does not cross-react with p53 phosphorylated at other sites.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser15 of human p53. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from MvILu cells treated with UV or hydroxyurea (20 mM) for the indicated times, using Phospho-p53 (Ser15) Antibody.
Western blot analysis of a p53 fusion protein, untreated or phosphorylated by DNA-PK, using Phospho-p53 (Ser15) Antibody (upper) and p53 Antibody #9282 (lower).
Western blot analysis of extracts from PC12 and HeLa cells treated with UV for the indicated times, using Phospho-p53 (Ser15) Antibody.
Confocal immunofluorescent analysis of HT-29 (p53++, left) and differentiated THP-1 cells (p53- -, right), UV-treated (top) or untreated (bottom), using Phospho-p53 (Ser15) Antibody (green). Actin filaments were labeled with DY-554 phalloidin (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT116 cells treated with UV (100 J/m2 followed by a 3 hour recovery) and either 5 μl of Phospho-p53 (Ser15) Antibody or 2 μl of 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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- Milczarek, G.J. et al. (1997) Life Sci. 60, 1-11.
- Shieh, S.Y. et al. (1997) Cell 91, 325-334.
- Chehab, N.H. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 13777-13782.
- Honda, R. et al. (1997) FEBS Lett. 420, 25-27.
- Tibbetts, R.S. et al. (1999) Genes Dev. 13, 152-157.
- Shieh, S.Y. et al. (1999) EMBO J. 18, 1815-1823.
- Hirao, A. et al. (2000) Science 287, 1824-1827.
- Hao, M. et al. (1996) J. Biol. Chem. 271, 29380-29385.
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- Lohrum, M. and Scheidtmann, K.H. (1996) Oncogene 13, 2527-2539.
- Knippschild, U. et al. (1997) Oncogene 15, 1727-1736.
- Oda, K. et al. (2000) Cell 102, 849-862.
- Ito, A. et al. (2001) EMBO J. 20, 1331-1340.
- Sakaguchi, K. et al. (1998) Genes Dev. 12, 2831-2841.
- Solomon, J.M. et al. (2006) Mol. Cell. Biol. 26, 28-38.
- Hirao, A. et al. (2000) Science 287, 1824-7. Applications: Western Blotting.
- Kruman, I.I. et al. (2000) J Neurosci 20, 6920-6. Applications: IC-IF.
- O'Driscoll, M. et al. (2003) Nat Genet 33, 497-501. Applications: IC-IF.
- Castedo, M. et al. (2004) Oncogene 23, 4353-61. Applications: Western Blotting, IC-IF.
- Castedo, M. et al. (2001) J. Exp. Med. 194, 1097-1110. Applications: IC-IF.
- Lim, D. S. et al. (2000) Nature 404, 613-617. Applications: Western Blotting.
- Lim, S. et al. (2009) Mol Cancer Res 7, 55-66. Applications: Western Blotting, IF-IC (In Cells).
- Raab, M. et al. (2011) Nat Commun 2, 395. Applications: Western Blotting.
- Jarvis, I.W. et al. (2013) Toxicol Appl Pharmacol 266, 408-18. Applications: Western Blotting.
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