Product Pathways - Cell Cycle / Checkpoint
Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific) #12571
|12571L||300 µl||---||In Stock||---|
|12571S||100 µl (10 western blots)||---||In Stock||---|
|12571P||40 µl||---||In Stock||---|
|12571||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Mouse, Rat||Endogenous||53||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Specificity / Sensitivity
Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific) recognizes endogenous levels of p53 protein only when phosphorylated at Ser15.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser15 of mouse p53 protein.
Western blot analysis of extracts from L-929 cells, untreated or treated with Etoposide #2200 (30 μM, 2 hours), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific). To characterize the phospo-specificity of the antibody, the blot was mock treated (-) or treated (+) with calf intestinal phosphatase (CIP).
Immunoprecipitation of phospho-p53 (Ser15) from etoposide-treated (30 μM, 2 hours) L-929 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific) (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific). A light-chain specific antibody was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded 4T1 metastatic tumors in mouse lung using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific). Note lack of staining in adjacent normal lung.
Immunohistochemical analysis of paraffin-embedded L-929 cell pellets, control (left) or treated with Etoposide #2200 (right), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific).
Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor, control (left) or λ phosphatase-treated (right), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific).
Flow cytometric analysis of L-929 cells, untreated (blue) or treated with Etoposide #2200 (green) (30 μM, 2 hours), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific). Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 conjugate) #4412 was used as a secondary antibody.
Confocal immunofluorescent analysis of L-929 cells, treated with Etoposide #2200 (left) or untreated (right), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific) (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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. (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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- Knippschild, U. et al. (1997) Oncogene 15, 1727-1736.
- Oda, K. et al. (2000) Cell 102, 849-862.
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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.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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