| 货号 | 12571L |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Mouse/Rat |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IHC-P/IF-IC/F |
| 目标/特异性 | Phospho-p53 (Ser15) (D4S1H) Rabbit mAb (Rodent Specific) recognizes endogenous levels of p53 protein only when phosphorylated at Ser15. |
| 使用方法 | WB(1:1000) IP (1:50) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | 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).p53抑癌蛋白在细胞对DNA损伤和其他基因组畸变过程中发挥了重要作用。激活p53将导致细胞周期停滞和DNA修复或凋亡（1）。体内实验发现p53可以在多个位点被磷酸化，体外实验也证明了几种不同的蛋白激酶都可以诱导这一过程（2,3）。DNA损伤会诱导p53第15和20位丝氨酸磷酸化，并减少p53和它的负调控因子，癌蛋白MDM2，的结合（4）。MDM2通过诱导泛素化和蛋白酶体降解以抑制p53（5,6）。ATM,ATR和DNA-PK都可以磷酸化p53的15和37位丝氨酸。磷酸化会影响MDM2与p53结合，从而促进p53聚集并激活p53对DNA损伤的反应（4,7）。Chk2和Chk1可以磷酸化p53的第20位丝氨酸，增强它的四聚体化，稳定性和活性（8,9）。p53在体内可以在392位丝氨酸处磷酸化（10,11），体外可以被CAK磷酸化（11）。392位丝氨酸磷酸化的p53在人类肿瘤中表达增加（12）并有报道显示该位点磷酸化会影响p53的生长抑制，结合DNA的功能和转录活性（10,13,14）。体内和体外实验都显示CK1δ and CK1ε能够磷酸化p53的第6和9位丝氨酸（16）。p53可以被p300和CBP乙酰转移酶乙酰化。p19（ARF）募集HDAC1复合物将抑制MDM2进而抑制p53去乙酰化使其稳定。在压力应激反应中乙酰化能够发挥促进p53蛋白富集的作用（17）。按随着DNA损伤，人p53蛋白的382位赖氨酸（小鼠是379位赖氨酸）在体内发生乙酰化以促进p53与DNA结合（18）。SIRT1蛋白，一种涉及到细胞衰老和DNA损伤反应的去乙酰基酶能够与p53结合，促使后者的去乙酰化（19）。 |
| 存放说明 | -20C |
| 计算分子量 | 53 |
| 参考文献 | 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. |
Flow cytometric analysis of L-929 cells, untreated (blue) or treated with Etoposide #2200 (green) (30 μM, 2 hours), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific). Anti-rabbit IgG (H+L), F(ab)2 fragment (Alexa Fluor® 488 conjugate) #4412 was used as a secondary antibody.未处理（蓝色）或经过Etoposide #2200处理（绿色）（30 μM，2小时）的L-929细胞，使用Phospho-p53 (Ser15) (D4S1H) XP®Rabbit mAb（Mouse Specific）进行流式分析。Anti-rabbit IgG (H+L), F(ab)2 fragment (Alexa Fluor® 488 conjugate) #4412作为二抗。 | |
Confocal immunofluorescent analysis of L-929 cells, treated with Etoposide #2200 (left) or untreated (right), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific) (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).经过Etoposide #2200处理（左）或未经处理（右）的L-929细胞，使用Phospho-p53 (Ser15) (D4S1H) XP®Rabbit mAb（Mouse Specific）（绿色）进行激光共聚焦免疫荧光分析。肌动蛋白丝使用DY-554鬼笔环肽标记（红色）。蓝色假色= DRAQ5® #4084 (fluorescent DNA dye). | |
Western blot analysis of extracts from L-929 cells, untreated or treated with Etoposide #2200 (30 μM, 2 hours), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific). To characterize the phospo-specificity of the antibody, the blot was mock treated (-) ot treated (+) with calf intestinal phosphatase (CIP).未处理或经过Etoposide #2200处理（30 μM，2小时）L-929细胞提取物，使用Phospho-p53 (Ser15) (D4S1H) XP®Rabbit mAb（Mouse Specific）进行western blot分析。为了区分该抗体的磷酸基团特异性，blot是未处理（-）或经过牛小肠碱性磷酸酶（CIP）处理（+）的。 | |
Immunohistochemical analysis of paraffin-embedded LL/2 syngeneic tumor, control (left) or λ phosphatase-treated (right), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific).使用Phospho-p53 (Ser15) (D4S1H) XP®Rabbit mAb (Mouse Specific)对对照（左）或λ磷酸酯酶处理（右）的石蜡包埋的LL/2同源性肿瘤进行免疫荧光分析。 | |
Immunohistochemical analysis of paraffin-embedded 4T1 metastatic tumors in mouse lung using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific). Note lack of staining in adjacent normal lung.使用Phospho-p53 (Ser15) (D4S1H) XP®Rabbit mAb (Mouse Specific)对石蜡包埋的4T1鼠肺转移瘤进行免疫荧光分析。注意：周边正常的肺组织未染色。 | |
Immunohistochemical analysis of paraffin-embedded L-929 cell pellets, control (left) or treated with Etoposide #2200 (right), using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific).使用Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific)对对照组（左）或Etoposide #2200处理后的石蜡包埋的L-929细胞进行免疫荧光分析。 | |
Immunoprecipitation of phospho-p53 (Ser15) from etoposide-treated (30 μM, 2 hours) L-929 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific) (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-p53 (Ser15) (D4S1H) XP® Rabbit mAb (Mouse Specific). A light-chain specific antibody was used as a secondary antibody.未处理（蓝色）或经过Etoposide #2200处理（绿色）（30 μM，2小时）的L-929细胞，使用Phospho-p53 (Ser15) (D4S1H) XP®Rabbit mAb (Mouse Specific)进行流式分析。Anti-rabbit IgG (H+L), F(ab)2 fragment (Alexa Fluor® 488 conjugate) #4412作为二抗。 |
