Product Pathways - DNA Damage
Phospho-p53 Antibody Sampler Kit #9919
|9919S||1 Kit (9 x 40 µl)||---||In Stock||---|
|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Phospho-p53 (Ser6) Antibody #9285||40 µl||W, IP||H, Mk||Hm||53||Rabbit|
|Phospho-p53 (Ser9) Antibody #9288||40 µl||W, IP||H, Mk||53||Rabbit|
|Phospho-p53 (Ser15) Antibody #9284||40 µl||W, IP, IF-IC, ChIP||H, M, R, Mk||Mi, B, Pg||53||Rabbit|
|p53 (7F5) Rabbit mAb #2527||40 µl||W, IHC-P, IF-IC, F, ChIP||H, Mk||53||Rabbit IgG|
|Phospho-p53 (Ser20) Antibody #9287||40 µl||W||H, Mk||53||Rabbit|
|Phospho-p53 (Ser37) Antibody #9289||40 µl||W, IF-IC, F||H, Mk||53||Rabbit|
|Phospho-p53 (Ser46) Antibody #2521||40 µl||W, IP, IF-IC, F||H, Mk||53||Rabbit|
|Phospho-p53 (Ser392) Antibody #9281||40 µl||W||H, M, Mi||53||Rabbit|
|Phospho-p53 (Thr81) Antibody #2676||40 µl||W, IHC-P||H, Mk||53||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry
Reactivity Key: H=Human, Mk=Monkey, M=Mouse, R=Rat, Mi=Mink
Western blot analysis of extracts from 293 and COS cells, using p53 (7F5) Rabbit mAb #2527.
Western blot analysis of extracts from HT29 cells, untreated, nocodazole-treated (50 ng/ml, 24h) or UV-treated (50mJ/cm2, 1hr), using Phospho-p53 (Thr81) Antibody #2676 (upper), p53 (1C12) Mouse mAb #2524 (middle), or Phospho-SAPK/JNK(T183/Y185) (98F2) Rabbit mAb #4671 (lower).
Western blot analysis of extracts from hydroxyurea (20 mM) treated Mv1Lu cells, using Phospho-p53 (Ser392) Antibody #9281.
Western blot analysis of extracts from UV or hydroxyurea (20 mM) treated Mv1Lu cells, using Phospho-p53 (Ser15) Antibody #9284.
Specificity and sensitivity of Phospho-p53 (Ser15) Antibody #9284. Western blot analysis of p53 fusion protein with and without DNA-PK phosphorylation, using Phospho-p53 (Ser15) Antibody (upper) and p53 Antibody #9282 (lower).
Western blot analysis of extracts from UV treated PC12 cells or HeLa cells, using Phospho-p53 (Ser15) Antibody #9284.
Western blot analysis of extracts from COS cells treated with UV or MMS, using Phospho-p53 (Ser6) Antibody #9285 or p53 Antibody #9282.
The Phospho-p53 Antibody Sampler Kit provides a fast and economical means of evaluating multiple phosphorylation sites of p53 protein. The kit contains enough primary and secondary antibodies to perform four Western blot experiments.
Specificity / Sensitivity
Phospho-p53 (Ser6), (Ser9), (Ser15), (Ser20), (Ser37), (Thr81), and (Ser392) Antibodies detect p53 only when phosphorylated at the indicated sites and do not cross-react with p53 phosphorylated at other sites.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser6, Ser9, Ser15, Ser20, Ser37, Thr81, and Ser392 of human p53. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a MBP-p53 fusion protein.
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).
- Levine, A.J. (1997) Cell 88, 323-331.
- Meek, D.W. (1994) Semin. Cancer Biol. 5, 203-210.
- 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.
- Lu, H. et al. (1997) Mol. Cell. Biol. 17, 5923-5934.
- Ullrich, S.J. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 5954-5958.
- Kohn, K.W. (1999) Mol. Biol. Cell 10, 2703-2734.
- 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.
- Lu, W.J. et al. (2011) Cell Death Differ 18, 1046-56. Applications: Western Blotting, IF-IC (In Cells).
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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.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.