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37909
p53 Antibody Sampler Kit

p53 Antibody Sampler Kit #37909

Western Blotting Image 1

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

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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 (Ser20) Antibody (upper) or p53 Antibody #9282 (lower).

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Western Blotting Image 3

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 4

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 5

Western blot analysis of extracts from MvILu cells treated with UV or hydroxyurea (20 mM) for the indicated times, using Phospho-p53 (Ser15) Antibody.

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Western Blotting Image 6

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 7

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 8

Western blot analysis of extracts from HeLa cells, untreated or treated with both trichostatin A #9950 (400 nM for 24 hours), and doxorubicin (0.5 µM for 24 hours) using Acetyl-p53 (Lys382) Antibody alone (A), antibody preincubated with a non-acetylated Lys382 peptide (B), or antibody preincubated with an acetylated Lys382 peptide (C).

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Western Blotting Image 9

Western blot analysis of extracts from NIH/3T3 (left) and MCF-7 (right) cells, untreated, doxorubicin-treated (0.5 μM, 24 hours) or doxorubicin and trichostatin A-treated (TSA, #9950, 400 nM, 24 hours), using Acetyl-p53 (Lys379) Antibody.

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Western Blotting Image 10

Western blot analysis of extracts from HT29 cells treated with nocodazole (50 ng/ml, 24h), calyculin A #9902 (100 nM, 10 min), or lambda Phosphatase NEB#P0753 (10,000 units/ml, 1 h), using Phospho-p53 (Ser33) Antibody (upper) or p53 (1C12) Mouse mAb #2524 (lower).

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IHC-P (paraffin) Image 11

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

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Western Blotting Image 12

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

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IP Image 13

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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Western Blotting Image 14

Western blot analysis of a p53 fusion protein, untreated or phosphorylated by DNA-PK, using Phospho-p53 (Ser15) Antibody (upper) and p53 Antibody #9282 (lower).

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Western Blotting Image 15

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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Western Blotting Image 16

Western blot analysis of extracts from HeLa cells, untreated, trichostatin A-treated #9950 (400 nM for 24 hours), doxorubicin-treated (0.5 µM for 24 hours), or both, using Acetyl-p53 (Lys382) Antibody (top) or p53 Antibody #2524 (bottom).

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IHC-P (paraffin) Image 17

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-p53 (Ser33) Antibody.

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IHC-P (paraffin) Image 18

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

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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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Western Blotting Image 20

Western blot analysis of extracts from PC12 cells treated with UV for the indicated times, using Phospho-p53 (Ser15) Antibody.

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IHC-P (paraffin) Image 21

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing nuclear localization, using Phospho-p53 (Ser33) Antibody.

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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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IF-IC Image 23

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 24

Immunohistochemical analysis of paraffin-embedded human lung dysplasia of alveolar cells, using Phospho-p53 (Ser33) Antibody in the presence of control peptide (left) or antigen specific peptide (right),

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Flow Cytometry Image 25

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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Chromatin IP Image 26

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 Phospho-p53 (Ser15) Antibody 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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IF-IC Image 27

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 28

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
p53 (7F5) Rabbit mAb 2527 20 µl
  • WB
  • IHC
  • IF
  • F
  • ChIP
H Mk 53 Rabbit IgG
Phospho-p53 (Ser20) Antibody 9287 20 µl
  • WB
H Mk 53 Rabbit 
Phospho-p53 (Ser392) Antibody 9281 20 µl
  • WB
H M Mi 53 Rabbit 
Phospho-p53 (Ser46) Antibody 2521 20 µl
  • WB
  • IP
  • IF
  • F
H Mk 53 Rabbit 
Phospho-p53 (Ser15) Antibody 9284 20 µl
  • WB
  • IP
  • ChIP
H M R Mk 53 Rabbit 
Phospho-p53 (Ser9) Antibody 9288 20 µl
  • WB
  • IP
H Mk 53 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 
Acetyl-p53 (Lys382) Antibody 2525 20 µl
  • WB
H 53 Rabbit 
Acetyl-p53 (Lys379) Antibody 2570 20 µl
  • WB
H M 53 Rabbit 
Phospho-p53 (Ser33) Antibody 2526 20 µl
  • WB
  • IHC
H Mk 53 Rabbit 

The p53 Antibody Sampler Kit provides an economical means of detecting p53 activity using modification-specific and control antibodies. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

p53 (7F5) Rabbit mAb detects endogenous levels of total p53 protein. This antibody binding has been mapped to the amino terminus region of human p53 protein.

Acetyl- and phospho-specific antibodies detect p53 only when phosphorylated or acetylated at the specified site.

p53 (7F5) Rabbit mAb is produced by immunizing animals with a full-length human p53 fusion protein. Polyclonal antibodies are produced by immunizing animals with a synthetic acetylated or phosphorylated peptide corresponding to residues surrounding Lys379 of mouse p53 (#2570) or to the specified site of human p53 (all others). Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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.
Entrez-Gene Id
7157
Swiss-Prot Acc.
P04637
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. 5,675,063.

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