Cell Signaling Technology

Product Pathways - DNA Damage

p53 (DO-7) Mouse mAb #48818

sc-126   sc-6243   sc-99  

No. Size Price
48818S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
48818 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 53 Mouse IgG2b
IHC-P 1:100
F 1:50
IF-IC 1:800
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

p53 (DO-7) Mouse mAb recognizes endogenous levels of total p53 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant human p53 protein expressed in E. coli.

IF-IC

IF-IC

Confocal immunofluorescent analysis of untreated HT-29 cells (top left), untreated Saos-2 cells (top right), untreated MCF7 cells (bottom left), and MCF7 cells treated with Doxorubicin #5927 (0.5 μM, 24 hr; bottom right), using p53 (DO-7) Mouse mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of H1299 cells (blue) and HT-29 cells (green) using p53 (DO-7) Mouse mAb. Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HT-29 (left) and Saos-2 (right) cells using p53 (DO-7) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using p53 (DO-7) Mouse mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT 116 cells treated with UV (100 J/m2 followed by a 3 hr recovery) and either 5 μl of p53 (DO-7) Mouse mAb or 2 μl of 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.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using p53 (DO-7) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from MCF7 cells, untreated (-) or treated with Doxorubicin #5927 (0.5 μM, 24 hr; +), using p53 (DO-7) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using p53 (DO-7) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using p53 (DO-7) Mouse mAb.

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.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalStain is a trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

Tween is a registered trademark of ICI Americas, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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