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65390
Phospho-p53 Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Phospho-p53 Antibody Sampler Kit #65390

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Phospho-p53 Antibody Sampler Kit: 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).

Phospho-p53 Antibody Sampler Kit: 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).

Phospho-p53 Antibody Sampler Kit: Image 3

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).

Phospho-p53 Antibody Sampler Kit: Image 4

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).

Phospho-p53 Antibody Sampler Kit: Image 5

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

Phospho-p53 Antibody Sampler Kit: Image 6

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).

Phospho-p53 Antibody Sampler Kit: Image 7

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.

Phospho-p53 Antibody Sampler Kit: Image 8

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.

Phospho-p53 Antibody Sampler Kit: Image 9

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).

Phospho-p53 Antibody Sampler Kit: Image 10

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).

Phospho-p53 Antibody Sampler Kit: Image 11

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).

Phospho-p53 Antibody Sampler Kit: Image 12

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).

Phospho-p53 Antibody Sampler Kit: Image 13

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).

Phospho-p53 Antibody Sampler Kit: Image 14

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

Phospho-p53 Antibody Sampler Kit: Image 15

Flow cytometric analysis of HT-29 cells using p53 (7F5) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Phospho-p53 Antibody Sampler Kit: Image 16

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).

Phospho-p53 Antibody Sampler Kit: Image 17

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.

Phospho-p53 Antibody Sampler Kit: Image 18

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).

Phospho-p53 Antibody Sampler Kit: Image 19

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

Phospho-p53 Antibody Sampler Kit: Image 20

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

Phospho-p53 Antibody Sampler Kit: Image 21

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

Phospho-p53 Antibody Sampler Kit: Image 22

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

Phospho-p53 Antibody Sampler Kit: Image 23

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.

Phospho-p53 Antibody Sampler Kit: Image 24

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

To Purchase # 65390T
Product # Size Price
65390T
1 Kit  (8 x 20 µl) N/A

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 (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 

Product Description

The Phospho-p53 Antibody Sampler Kit provides a fast and economical means of evaluating multiple phosphorylation sites of p53 protein. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Phospho-p53 (Ser6), (Ser9), (Ser15), (Ser20), (Ser46), (Thr81), and (Ser392) Antibodies detect p53 only when phosphorylated at the indicated sites and do not cross-react with p53 phosphorylated at other sites. p53 (7F5) Rabbit mAb detects endogenous levels of total p53 protein.

Source / Purification

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.

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, 323-31.
  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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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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