CST, 2524L, p53 (1C12) Mouse Monoclonal Antibody

Monoclonal Antibody for studying p53. Validated for WB,IP,IF,F,ChIP. Available in 3 sizes. Highly specific and rigorously validated in-house, p53 (1C12) Mouse Monoclonal Antibody (CST #2524) is ready to ship. Product Usage Information For optimal ChIP results, use 2.5 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. Western Blotting: 1:1000 Immunoprecipitation: 1:500 Immunofluorescence (Immunocytochemistry): 1:3200 - 1:12800 Flow Cytometry (Fixed/Permeabilized): 1:800 - 1:3200 Chromatin IP: 1:200 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. For a carrier free (BSA and azide free) version of this product see product # 15755 . Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP Specificity / Sensitivity p53 (1C12) Mouse Monoclonal Antibody detects endogenous levels of total p53 protein. Species Reactivity: Human, Mouse, Rat, Hamster, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser20 of human p53. Background 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 and by several different protein kinases (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 (10,11) and by CAK (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 and (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) 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). Alternate Names Antigen NY-CO-13; BCC7; BMFS5; Cellular tumor antigen p53; FLJ92943; LFS1; mutant tumor protein 53; P53; p53 antigen; p53 transformation suppressor; p53 tumor suppressor; Phosphoprotein p53; TP53; transformation-related protein 53; TRP53; tumor protein 53; tumor protein p53; Tumor suppressor p53; tumor supressor p53 Specification REACTIVITY: H M R Hm Mk SENSITIVITY: Endogenous MW (kDa): 53 Source/Isotype: Mouse IgG1