Cell Signaling Technology

Product Pathways - DNA Damage

p53 (1C12) Mouse mAb #2524

sc-126   sc-6243   sc-99  

No. Size Price
2524S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
2524T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价
2524 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Hamster,Monkey, Endogenous 53 Mouse IgG1
IP 1:500
F 1:3200
IF-IC 1:4000
ChIP 1:200

Species cross-reactivity is determined by western blot.

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

Specificity / Sensitivity

p53 (1C12) Mouse mAb detects endogenous levels of total p53 protein.p53(1C12)Mouse mAb能够检测内源性p53总蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser20 of human p53.该单克隆抗体是由合成的人源的针对p53蛋白丝氨酸(20位)的肽段免疫动物生产的。

Western Blotting

Western Blotting

Western blot analysis of extracts from A431, COS, NBT-II and JB6 cells, untreated or UV-treated, using p53 (1C12) Mouse mAb.Western blot 方法检测未处理或紫外处理的A431, COS, NBT-II 和 JB6 细胞提取物,使用的抗体为p53 (1C12) Mouse mAb。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HT-29 cells using p53 (1C12) Mouse mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).激光共聚焦免疫荧光方法检测HT-29细胞,使用的抗体为p53 (1C12) Mouse mAb,呈绿色。肌动蛋白纤维由DY-554鬼笔环肽标记,呈红色。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p53 siRNA II (+), using p53 (1C12) Mouse mAb and β-Actin (13E5) Rabbit mAb #4970. p53 (1C12) Mouse mAb confirms silencing of p53 expression, while the β-Actin (13E5) Rabbit mAb is used to control for loading and specificity of p53 siRNA.Western blot方法检测Hela细胞提取物,分别转染100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) 或SignalSilence® p53 siRNA II (+),使用的抗体为p53 (1C12) Mouse mAb 和β-Actin (13E5) Rabbit mAb #4970。与对照抗体β-Actin (13E5) Rabbit mAb相比,p53 (1C12) Mouse mAb抗体证明了p53 siRNA能够特异性沉默p53蛋白表达。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p53 siRNA I (+), using p53 (1C12) Mouse mAb and p42 MAPK (Erk2) Antibody #9108. p53 (1C12) Mouse mAb confirms silencing of p53 expression, while the p42 MAPK (Erk2) Antibody is used to control for loading and specificity of p53 siRNA.Western blot方法检测Hela细胞提取物,分别转染100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) 或 SignalSilence® p53 siRNA I (+),使用的抗体为p53 (1C12) Mouse mAb 和 p42 MAPK (Erk2) Antibody #9108。与对照抗体p42 MAPK (Erk2) Antibody相比,p53 (1C12) Mouse mAb抗体证明了p53 siRNA能够特异性沉默p53蛋白表达。

Chromatin IP

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 5 μl of p53 (1C12) 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.采用来自4 x 106 HCT116细胞的交叉结合染色质进行免疫共沉淀实验,细胞通过紫外线处理(100 J/m2,恢复3小时)和5 μl p53 (1C12) Mouse mAb或者2 μl Normal Rabbit IgG #2729,使用的试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。随后通过实时PCR定量富集的DNA,使用的引物为SimpleChIP® Human CDKN1A Promoter Primers #6449,人 MDM2 内含子 2 primers,和 SimpleChIP® Human α Satellite Repeat Primers #4486。每个样品的免疫沉淀DNA的量以相对于输入染色质总量(相当于1)的信号来表示

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). p53肿瘤抑制蛋白在细胞响应DNA损伤和其它基因组异常的过程中发挥重要作用。p53的激活能够引起细胞周期捕获和DNA修复或细胞凋亡(1)。离体或者在体情况下,p53可以在多个位点被几个蛋白激酶进行磷酸化(2,3)。DNA损伤能够诱导p53的第15位和20位丝氨酸磷酸化,导致p53和其负调节子-癌蛋白MDM2的相互作用减弱(4)。MDM2通过靶向p53促进其泛素化和蛋白酶体降解抑制其累积。P53的第15位和37位丝氨酸可以被ATM,ATR,和DNA-PK磷酸化(5,6)。P53的磷酸化削弱了MDM2的结合,从而促进了它的激活和累积以响应DNA损伤(4,7)。Chk2和Chk1能够磷酸化p53的第20位丝氨酸,增强其四聚化、稳定性和活性(8,9)。p53的第392位丝氨酸可以发生在体磷酸化(10,11)和CAK介导的离体磷酸化(11)。人类肿瘤中p53的第392位丝氨酸磷酸化增加(12)且有报道认为该过程能够影响生长抑制剂的功能、DNA结合和p53的转录激活(10,13,14)。P53的第6位和9位丝氨酸离体或者在体均可以被CK1δ和CK1ε磷酸化(13,15)。P53的第46位丝氨酸磷酸化能够调节p53诱导细胞凋亡的能力(16)。p300和CBP乙酰转移酶能够介导p53的乙酰化。去乙酰化抑制可以阻止MDM2招募p19 (ARF)介导的HDAC1复合体从而稳定p53。乙酰化似乎对于应激反应中p53蛋白的累积起着正向的作用(17)。DNA损伤后,人类p53的第382位赖氨酸(在小鼠为379位赖氨酸)发生在体乙酰化,促进p53-DNA的结合(18)。P53通过与SIRT1蛋白(一种参与细胞衰老和DNA损伤应激的去乙酰酶)相互作用发生去乙酰化(19)。

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