Product Pathways - DNA Damage
p53 (1C12) Mouse mAb #2524
|2524S||100 µl (10 western blots)||---||In Stock||---|
|2524||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat, Monkey||Endogenous||53||Mouse IgG1|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP
Specificity / Sensitivity
p53 (1C12) Mouse mAb detects endogenous levels of total p53 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser20 of human p53.
Western blot analysis of extracts from A431, COS, NBT-II and JB6 cells, untreated or UV-treated, using p53 (1C12) Mouse mAb.
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p53 siRNA I (+), using p53 (1C12) Mouse mAb and p42 MAPK (Erk2) Antibody #9108. p53 (1C12) Mouse mAb confirms silencing of p53 expression, while the p42 MAPK (Erk2) Antibody is used to control for loading and specificity of p53 siRNA.
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p53 siRNA II (+), using p53 (1C12) Mouse mAb and β-Actin (13E5) Rabbit mAb #4970. p53 (1C12) Mouse mAb confirms silencing of p53 expression, while the β-Actin (13E5) Rabbit mAb is used to control for loading and specificity of p53 siRNA.
Confocal immunofluorescent analysis of HT-29 cells using p53 (1C12) Mouse mAb (green). Actin filaments have been 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 p53 (1C12) Mouse mAb 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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- Sakaguchi, K. et al. (1998) Genes Dev. 12, 2831-2841.
- Solomon, J.M. et al. (2006) Mol. Cell. Biol. 26, 28-38.
- Lim, S. et al. (2009) Mol Cancer Res 7, 55-66. Applications: Western Blotting.
- Textor, S. et al. (2011) Cancer Res , . Applications: Chromatin IP.
- Zimnik, S. et al. (2009) Nucleic Acids Res 37, e30. Applications: Western Blotting.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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