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Phospho-p53 Antibody Sampler Kit #9919

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
Phospho-p53 (Ser6) Antibody #9285	40 µl	W IP IC	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	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	53	Rabbit
Phospho-p53 (Thr81) Antibody #2676	40 µl	W IHC-P IF-IC	H Mk	53	Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  IC=Immunocytochemistry  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

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.

Description

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.

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.

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 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).

1. Levine, A.J. (1997) Cell 88, 323-331.
2. Meek, D.W. (1994) Semin. Cancer Biol. 5, 203-210.
3. Milczarek, G.J. et al. (1997) Life Sci. 60, 1-11.
4. Shieh, S.Y. et al. (1997) Cell 91, 325-334.
5. Chehab, N.H. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 13777-13782.
6. Honda, R. et al. (1997) FEBS Lett. 420, 25-27.
7. Tibbetts, R.S. et al. (1999) Genes Dev. 13, 152-157.
8. Shieh, S.Y. et al. (1999) EMBO J. 18, 1815-1823.
9. Hirao, A. et al. (2000) Science 287, 1824-1827.
10. Hao, M. et al. (1996) J. Biol. Chem. 271, 29380-29385.
11. Lu, H. et al. (1997) Mol. Cell. Biol. 17, 5923-5934.
12. Ullrich, S.J. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 5954-5958.
13. Kohn, K.W. (1999) Mol. Biol. Cell 10, 2703-2734.
14. Lohrum, M. and Scheidtmann, K.H. (1996) Oncogene 13, 2527-2539.
15. Knippschild, U. et al. (1997) Oncogene 15, 1727-1736.
16. Oda, K. et al. (2000) Cell 102, 849-862.
17. Ito, A. et al. (2001) EMBO J. 20, 1331-1340.
18. Sakaguchi, K. et al. (1998) Genes Dev. 12, 2831-2841.
19. Solomon, J.M. et al. (2006) Mol. Cell. Biol. 26, 28-38.

Application References

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Protocols

2521:

Flow, Immunofluorescence, Immunoprecipitation, Western Blotting

2527:

ChIP Agarose, ChIP Magnetic, Flow, IHC / Paraffin, Immunofluorescence, Western Blotting

2676:

IHC / Paraffin, Western Blotting

9281:

Western Blotting

9284:

ChIP Agarose, ChIP Magnetic, Immunofluorescence, Immunoprecipitation, Western Blotting

9285:

Immunoprecipitation, Western Blotting

9287:

Western Blotting

9288:

Immunoprecipitation, Western Blotting

9289:

Flow, Immunofluorescence, Western Blotting

Companion Products

• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7076 Anti-mouse IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9917 Cell Cycle/Checkpoint Antibody Sampler Kit
• 9931 Phospho-Chk1/2 Antibody Sampler Kit
• 9947 DNA Damage Antibody Sampler Kit
• 8342 UV Induced DNA Damage Response Antibody Sampler Kit
• 8344 MRN Complex Antibody Sampler Kit
• 9786 Mismatch Repair Antibody Sampler Kit

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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.