DNA Damage
- DNA Damage
PhosphoSitePlus®：
p53
- 推奨プロトコール
p53 (7F5) Rabbit mAb |
イイネ！(3) | 
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| CSTコード | 包装 | 希望納入価格 (円) |
国内在庫  2012年2月9日15時20分 現在 |
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| #2527S | 100 μL | 46,000 | ログインすると国内在庫状況がご確認いただけます。
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2527 の推奨プロトコール 最適な結果を得るために：Cell Signaling Technology (CST) 社は、各製品の推奨プロトコールを使用することを強くお薦めいたします。 注：各製品に最適化されたプロトコールをリンクしています。 |
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下記ステップについては、データシートの右側もあわせてご参照ください。
IHC-P: 抗体希釈液 / 抗原賦活化
| 用途 (希釈倍率) | |
|---|---|
| ウェスタンブロッティング (1:1,000)、免疫組織染色 (パラフィン) (1:40)、免疫蛍光細胞染色 (IF-IC) (1:400)、フローサイトメトリー (1:100)、ChIP (1:50) |
| 種交差性 | |
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| ヒト、サル |
| 特異性・感度 | |
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| 内在性レベルのp53 タンパク質を検出します。p53 タンパク質のN末端領域に結合します。 |
| 検出タンパク質の分子量 | |
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| 53 kDa |
| 使用抗原 | |
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| ヒトの全長p53 融合タンパク質 |
| 抗体の由来 | |
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| ウサギ |
| 貯法 | |
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| -20℃ |
| 社内データ |
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Western Blotting
Western blot analysis of extracts from 293 and COS cells, using p53 (7F5) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using p53 (7F5) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using p53 (7F5) Rabbit mAb.
IHC-P (paraffin)
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.
Flow Cytometry
Flow cytometric analysis of HT-29 cells using p53 (7F5) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
IF-IC
Confocal Immunofluorescent analysis of HT-29 cells using p53 (7F5) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HCT116 cells treated with UV (100 J/m2 followed by a 3 hour recovery) and either 10 μl of p53 (7F5) Rabbit 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.
| バックグラウンド |
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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).
- Levine, A.J. (1997) Cell 88, 323-331.
- Meek, D.W. (1994) Semin. Cancer Biol. 5, 203-210.
- Milczarek, G.J. et al. (1997) Life Sci. 60, 1-11.
- Shieh, S.Y. et al. (1997) Cell 91, 325-334.
- Chehab, N.H. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 13777-13782.
- Honda, R. et al. (1997) FEBS Lett. 420, 25-27.
- Tibbetts, R.S. et al. (1999) Genes Dev. 13, 152-157.
- Shieh, S.Y. et al. (1999) EMBO J. 18, 1815-1823.
- Hirao, A. et al. (2000) Science 287, 1824-1827.
- Hao, M. et al. (1996) J. Biol. Chem. 271, 29380-29385.
- Lu, H. et al. (1997) Mol. Cell. Biol. 17, 5923-5934.
- Ullrich, S.J. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 5954-5958.
- Kohn, K.W. (1999) Mol. Biol. Cell 10, 2703-2734.
- Lohrum, M. and Scheidtmann, K.H. (1996) Oncogene 13, 2527-2539.
- Knippschild, U. et al. (1997) Oncogene 15, 1727-1736.
- Oda, K. et al. (2000) Cell 102, 849-862.
- Ito, A. et al. (2001) EMBO J. 20, 1331-1340.
- Sakaguchi, K. et al. (1998) Genes Dev. 12, 2831-2841.
- Solomon, J.M. et al. (2006) Mol. Cell. Biol. 26, 28-38.
| 使用例 | |
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本製品は試験研究用です。