Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry)

Specificity / Sensitivity

53BP1 Antibody detects endogenous levels of total 53BP1 protein independent of phosphorylation. 53BP1抗体检测内源性独立于磷酸化的53BP1总蛋白水平。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the center of human 53BP1. Antibodies are purified by protein A and peptide affinity chromatography. 该多克隆抗体由合成的人源的针对53BP1蛋白近中心区域残基的肽段免疫动物，利用A蛋白和多肽亲和层析的方法纯化获得。

Western Blotting

Western blot analysis of extracts from HT29 cells, using 53BP1 Antibody. Western blot 方法检测HT29细胞提取物中53BP1蛋白水平，使用的抗体为53BP1 。

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using 53BP1 Antibody. 免疫组织化学方法检测石蜡包埋的人乳腺癌组织，图示为核定位。使用的抗体为53BP1 。

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma, using 53BP1 Antibody. 免疫组织化学方法检测石蜡包埋的人肾细胞癌组织，使用的抗体为53BP1 。

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using 53BP1 Antibody in the presence of control peptide (left) or antigen-specific peptide (right). 免疫组织化学方法检测石蜡包埋的人结肠癌组织，使用的抗体为53BP1 。左图是对照组，右图是抗原特异性肽段组。

IF-IC

Confocal immunofluorescent analysis of HeLa cells using 53BP1 Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). 激光共聚焦荧光法检测HeLa 细胞，使用的抗体为53BP1(绿色)。肌动蛋白纤维采用Alexa Fluor® 555鬼笔环肽标记(红色)。

Background

p53-binding protein 1 (53BP1) was originally identified as a p53 binding partner that could enhance the transcriptional activity of p53 (1,2). 53BP1 consists of two BRCA1 carboxy terminal (BRCT) domains that allow for binding to p53 and a separate domain responsible for binding to phosphorylated histone H2A.X (3). 53BP1 rapidly translocates to nuclear foci following treatment of cells with ionizing radiation (IR) or radiomimetic agents that cause DNA double strand breaks (DSBs) (4,5). Because of this localization to DSBs and homology to the yeast protein Rad9, a role for 53BP1 in DSB repair has been proposed. Recruitment of 53BP1 to sites of DNA damage has been demonstrated to be independent of ATM, NBS1, and DNA-PK (4) and retention of 53BP1 at DNA breaks requires phosphorylated H2A.X (6). In cells lacking 53BP1, phosphorylation of ATM substrates is reduced, suggesting that 53BP1 is upstream of ATM (7). In response to IR, phosphorylation of 53BP1 at serines 6, 25, 29, and 784 by ATM has been demonstrated, but phosphorylation at these sites is not required for localization of 53BP1 to sites of DSBs (6). p53结合蛋白1（p53 binding protein 1，53BPl）最初被认为是一种p53的结合蛋白，其功能是增强p53基因的转录活性。53BP1通过其两个BRCA1羧基末端（BRCT）结构域与p53基因结合，通过另外一个独立的结构域与磷酸化组蛋白H2A.X结合。电离辐射（IR）或者辐射类物预处理细胞后，可引起DNA双链断裂（DSBs），53BP1蛋白迅速转移到核内。鉴于DNA断裂后53BP1的定位改变及其与酵母蛋白Rad9的同源性，人们提出53BP1可能在DNA双链断裂后的修复过程中起到一定的作用。53BP1聚集到DNA损伤处的过程不依赖于ATM,NBS1,和DNA-PK，但是其在DNA断裂处的滞留依赖于磷酸化H2A.X。在53BP1表达缺失的细胞中，ATM底物的磷酸化程度降低，提示53BP1在ATM上游起作用。电离刺激后，53BP1在丝氨酸6,25,29以及784位被ATM磷酸化，但是这些位点的磷酸化并不影响53BP1在DNA双链断裂处的聚集。

12630 SignalFire™ Plus ECL Reagent

12757 SignalFire™ Elite ECL Reagent

2524 p53 (1C12) Mouse mAb

2577 Phospho-Histone H2A.X (Ser139) Antibody

2661 Phospho-Chk2 (Thr68) Antibody

2674 Phospho-53BP1 (Ser25/29) Antibody

2675 Phospho-53BP1 (Ser1778) Antibody

3002 p95/NBS1 Antibody

3428 Phospho-53BP1 (Thr543) Antibody

6209 Phospho-53BP1 (Ser1618) (D4H11) Rabbit mAb

6883 SignalFire™ ECL Reagent

7074 Anti-rabbit IgG, HRP-linked Antibody

7720 Prestained Protein Marker, Broad Range (Premixed Format)

7727 Biotinylated Protein Ladder Detection Pack