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9919
Phospho-p53 Antibody Sampler Kit

Phospho-p53 Antibody Sampler Kit #9919

Western Blotting Image 1

Western blot analysis of extracts from COS cells treated with UV or MMS for the indicated times, using Phospho-p53 (Ser6) Antibody (upper) or p53 Antibody #9282 (lower).

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Western Blotting Image 2

Western blot analysis of extracts from COS cells treated with UV or MMS for the indicated times, using Phospho-p53 (Ser9) Antibody (upper) or p53 Antibody #9282 (lower).

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Western Blotting Image 3

Western blot analysis of extracts from HT29 cells, untreated or UV-treated (100 mJ/cm2, 1 hr), using Phospho-p53 (Ser15) (16G8) Mouse mAb (upper) or p53 (DO-7) Mouse mAb #48818 (lower).

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Western Blotting Image 4

Western blot analysis of extracts from MCF-7 cells treated with doxorubicin (0.5 uM, 24 h) or doxorubicin plus lambda Phosphatase NEB#P0753 (10,000 Units/ml, 1h), using Phospho-p53 (Thr18) Antibody (upper) or p53 Antibody #9282 (lower).

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Western Blotting Image 5

Western blot analysis of extracts from COS cells treated with UV or MMS for the indicated times, using Phospho-p53 (Ser20) Antibody (upper) or p53 Antibody #9282 (lower).

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Western Blotting Image 6

Western blot analysis of extracts from MCF-7 cells treated with etoposide for the indicated times, using Phospho-p53 (Ser46) Antibody.

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Western Blotting Image 7

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 (upper), p53 (1C12) Mouse mAb #2524 (middle), or Phospho-SAPK/JNK(T183/Y185) (98F2) Rabbit mAb #4671 (lower).

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Western Blotting Image 8

Western blot analysis of increasing amounts of a p53 fusion protein, untreated or phosphorylated by CKII, using Phospho-p53 (Ser392) Antibody (upper) or p53 Antibody #9282 (lower).

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Western Blotting Image 9

Western blot analysis of extracts from 293 and COS cells, using p53 (7F5) Rabbit mAb.

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Western Blotting Image 10

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Western Blotting Image 11

Western blot analysis of a p53 fusion protein, untreated or phosphorylated by CK1 or CK2, using Phospho-p53 (Ser6) Antibody (upper) or p53 Antibody #9282 (lower).

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Western Blotting Image 12

Western blot analysis of a p53 fusion protein, untreated or phosphorylated by CK1 or CK2, using Phospho-p53 (Ser9) Antibody (upper) or p53 Antibody #9282 (lower).

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Flow Cytometry Image 13

Flow cytometric analysis of HT-29 cells, untreated (blue) or UV-treated (green), using Phospho-p53 (Ser15) (16G8) Mouse mAb compared to a nonspecific negative control antibody (red).

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IP Image 14

Immunoprecipitation of extracts from MCF-7 cells treated with etoposide under nondenaturing conditions, using Phospho-p53 (Ser46) Antibody, followed by Western blot analysis using a monoclonal p53 antibody.

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IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded HT-29 cells, untreated (left) or nocodazole-treated (right), using Phospho-p53 (Thr81) Antibody in the presence of control peptide (lower) or antigen-specific peptide (upper).

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Western Blotting Image 16

Western blot analysis of extracts from Mv1Lu cells, untreated or hydroxyurea-treated (20 mM), using Phospho-p53 (Ser392) Antibody.

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IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using p53 (7F5) Rabbit mAb.

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IF-IC Image 18

Confocal immunofluorescent analysis of HT-29 cells, untreated (left) or UV-treated (right), using Phospho-p53 (Ser15) (16G8) Mouse mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

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Flow Cytometry Image 19

Flow cytometric analysis of HT-29 cells, untreated (blue) or UV-treated (green), using Phospho-p53 (Ser46) Antibody.

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IHC-P (paraffin) Image 20

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using p53 (7F5) Rabbit mAb.

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IF-IC Image 21

Confocal immunofluorescent analysis of MCF-7 cells, untreated (left) or etoposide-treated (right), using Phospho-p53 (Ser46) Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded HT-29 (left) and SaOs-2 (right) cells, using p53 (7F5) Rabbit mAb. Note the lack of staining in p53-negative SaOs-2 cells.

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Flow Cytometry Image 23

Flow cytometric analysis of HT-29 cells using p53 (7F5) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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IF-IC Image 24

Confocal Immunofluorescent analysis of HT-29 cells using p53 (7F5) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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Chromatin IP Image 25

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells treated with UV (100 J/m2 followed by a 3 hour recovery) and either p53 (7F5) Rabbit mAb or 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.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-p53 (Ser6) Antibody 9285 20 µl
  • WB
  • IP
H Mk 53 Rabbit 
Phospho-p53 (Ser9) Antibody 9288 20 µl
  • WB
  • IP
H Mk 53 Rabbit 
Phospho-p53 (Ser15) (16G8) Mouse mAb 9286 20 µl
  • WB
  • IF
  • F
H 53 Mouse IgG1
Phospho-p53 (Thr18) Antibody 2529 20 µl
  • WB
H 53 Rabbit 
Phospho-p53 (Ser20) Antibody 9287 20 µl
  • WB
H Mk 53 Rabbit 
Phospho-p53 (Ser46) Antibody 2521 20 µl
  • WB
  • IP
  • IF
  • F
H Mk 53 Rabbit 
Phospho-p53 (Thr81) Antibody 2676 20 µl
  • WB
  • IHC
H Mk 53 Rabbit 
Phospho-p53 (Ser392) Antibody 9281 20 µl
  • WB
H M Mi 53 Rabbit 
p53 (7F5) Rabbit mAb 2527 20 µl
  • WB
  • IHC
  • IF
  • F
  • ChIP
H Mk 53 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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 two Western blot experiments.

Phospho-p53 (Ser6), (Ser9), (Ser15), (Ser20), (Thr18), (Thr81), and (Ser392) Antibodies detect p53 only when phosphorylated at the indicated sites and do not cross-react with p53 phosphorylated at other sites.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide and are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with recombinant human proteins or synthetic peptides.

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, 323-31.
  2. Meek, D.W. (1994) Semin Cancer Biol 5, 203-10.
  3. Milczarek, G.J. et al. (1997) Life Sci 60, 1-11.
  4. Shieh, S.Y. et al. (1997) Cell 91, 325-34.
  5. Chehab, N.H. et al. (1999) Proc Natl Acad Sci U S A 96, 13777-82.
  6. Honda, R. et al. (1997) FEBS Lett 420, 25-7.
  7. Tibbetts, R.S. et al. (1999) Genes Dev 13, 152-7.
  8. Shieh, S.Y. et al. (1999) EMBO J 18, 1815-23.
  9. Hirao, A. et al. (2000) Science 287, 1824-7.
  10. Hao, M. et al. (1996) J Biol Chem 271, 29380-5.
  11. Lu, H. et al. (1997) Mol Cell Biol 17, 5923-34.
  12. Ullrich, S.J. et al. (1993) Proc Natl Acad Sci U S A 90, 5954-8.
  13. Kohn, K.W. (1999) Mol Biol Cell 10, 2703-34.
  14. Lohrum, M. and Scheidtmann, K.H. (1996) Oncogene 13, 2527-39.
  15. Knippschild, U. et al. (1997) Oncogene 15, 1727-36.
  16. Oda, K. et al. (2000) Cell 102, 849-62.
  17. Ito, A. et al. (2001) EMBO J 20, 1331-40.
  18. Sakaguchi, K. et al. (1998) Genes Dev 12, 2831-41.
  19. Solomon, J.M. et al. (2006) Mol Cell Biol 26, 28-38.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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