Cell Signaling Technology

Product Pathways - DNA Damage

Double Strand Breaks (DSB) Repair Antibody Sampler Kit #9653

atm  

REACTIVITY

No. Size Price
9653T 1 Kit ( 9 x 20 µl ) ¥7,245.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Ku80 (C48E7) Rabbit mAb #2180 20 µl W,IP,IHC-P,IHC-F,IF-IC, H,Mk, 86 Rabbit IgG
XLF Antibody #2854 20 µl W,IP, H, 39 Rabbit
ATM (D2E2) Rabbit mAb #2873 20 µl W, H,M, 350 Rabbit
Phospho-p95/NBS1 (Ser343) Antibody #3001 20 µl W, H,M,Mi, 95 Rabbit
Rad50 Antibody #3427 20 µl W, H,Mk, 153 Rabbit
DNA-PKcs Antibody #4602 20 µl W, H, 450 Rabbit
Mre11 (31H4) Rabbit mAb #4847 20 µl W,IP,IHC-P,IHC-F, H, 81 Rabbit IgG
Phospho-ATM (Ser1981) (D6H9) Rabbit mAb #5883 20 µl W, H, Mk,B,Pg,Hr, 350 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W, Goat
Phospho-BRCA1 (Ser1524) Antibody #9009 20 µl W, H, 220 Rabbit

Specificity / Sensitivity

Each antibody in the Double Strand Breaks (DSB) Repair Antibody Sampler Kit detects endogenous levels of its respective protein and does not cross-react with other family members. Activation state antibodies only detect their target proteins when modified at the indicated site.Double Strand Breaks (DSB) Repair Antibody Sampler Kit中的每个抗体都能够检测其内源性靶蛋白,且不与家族其他成员发生交叉反应。激活状态的抗体仅能检测特定位点修饰的靶蛋白。

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Lys496 of human Mre11A and the carboxy terminus of human Ku80. Activation state monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser1981 of human ATM. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the carboxy-terminus of human DNA-PKcs, the amino terminus of human Rad50, the central sequence of human Rad52, and the carboxy terminus of human XLF. Activation state polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser1524 of human BRCA1 and surrounding Ser343 of human p95/NBS1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.单克隆抗体是由合成的人源的针对Mre11A蛋白赖氨酸(496位)的肽段 和Ku80蛋白羧基末端肽段免疫动物生产的。激活状态的单克隆抗体是由合成的人源的针对ATM蛋白丝氨酸(1981位)的磷酸化肽段免疫动物生产的。多克隆抗体是由人源的针对DNA-PKcs蛋白的羧基末端肽段、Rad50的氨基末端肽段,Rad52蛋白的中间肽段及XLF蛋白的羧基末端肽段免疫动物生产的。激活状态的多克隆抗体是由合成的人源的针对BRCA1蛋白丝氨酸(1524位)的磷酸化肽段及p95/NBS1蛋白的丝氨酸(343位)的磷酸化肽段免疫动物,采用蛋白A和多肽亲和层析技术纯化生产的。

Description

The Double Strand Breaks (DSB) Repair Antibody Sampler Kit provides an economical means to investigate repair of double-strand DNA breaks within the cell. The kit contains primary and secondary antibodies to perform four western blots with each antibody.Double Strand Breaks (DSB) Repair Antibody Sampler Kit能够经济地检测细胞内双链DNA断裂修复。该试剂盒内含足够4次western blot实验的一抗和二抗。

Western Blotting

Western Blotting

Western blot analysis of extracts from Mv1Lu cells treated with UV or hydroxyurea (HU) for the indicated times, using Phospho-p95/NBS1 (Ser343) Antibody.

Western Blotting

Western Blotting

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

Western Blotting

Western Blotting

Western blot analysis of extracts from M059K (DNA-PK wildtype) and M059J (DNA-PK deficient) cells, using DNA-PK Antibody.

IF-IC

IF-IC

Immunofluorescent analysis of M059J cells, showing localization primarily in the nucleoli, using Rad52 Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from K562 cells, using Rad52 Antibody.

Western Blotting

Western Blotting

Western blot analysis of untreated and UV-treated (50 mJ/cm2,30 min) HeLa cells and HT-1376 cells, using Phospho-BRCA1 (Ser1524) Antibody (upper) and BRCA1 Antibody #9010 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and K562 cells, using Mre11 Rabbit (31H4) mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded lung carcinoma, using Mre11 (31H4) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Mre11 (31H4) Rabbit mAb in the presence of control peptide (left) or Mre11 Blocking Peptide #1035 (right).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of K562 cells, using Mre11 (31H4) Rb mAb (blue) compared to a nonspecific negative control antibody (red).

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat and K562 cells, using RAD50 Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Ku80 (C48E7) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Ku80 (C48E7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

Western Blotting

Western Blotting

Western blot analysis of cell extracts from HeLa, A549 and COS cells using Ku80 (C48E7) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human glioblastoma using Ku80 (C48E7) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human GIST using Ku80 (C48E7) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human melanoma using Ku80 (C48E7) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts of various cell lines using XLF Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts of HeLa, NCCIT and PYS2 cells using ATM (D2E2) Rabbit mAb.

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen SKOV-3 xenograft using Ku80 (C48E7) Rabbit mAb.

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen SKOV-3 xenograft using Mre11 (31H4) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, A549 and COS cells using Ku80 (C48E7) Rabbit mAb #2180.Western blot方法检测 HeLa, A549 和COS细胞提取物,使用的抗体为Ku80 (C48E7) Rabbit mAb #2180。

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using XLF Antibody #2854.Western blot方法检测不同细胞提取物,使用的抗体为XLF Antibody #2854.

Western Blotting

Western Blotting

Western blot analysis of extracts from K-562 cells using Rad52 Antibody #3425.Western blot检测k-562细胞提取物,使用的抗体为Rad52 Antibody #3425。

Western Blotting

Western Blotting

Western blot analysis of extracts from Mv 1 Lu cells, untreated, UV-treated, or hydroxyurea (HU) treated for the indicated times, using Phospho-p95/NBS1 (Ser343) Antibody #3001.Western blot方法检测未处理、紫外处理和羟基脲处理不同时间的Mv1Lu细胞提取物,使用的抗体为Phospho-p95/NBS1 (Ser343) Antibody #3001。

Western Blotting

Western Blotting

Western blot analysis of HeLa and HT-1376 cells, untreated or UV-treated (50 mJ/cm2 for 30 min), using Phospho-BRCA1 (Ser1524) Antibody #9009 (upper) and BRCA1 Antibody #9010 (lower).Western blot 方法检测未处理和紫外处理(50 mJ/cm2 )30分钟的HeLa 和HT-1376 细胞,使用的抗体为 Phospho-BRCA1 (Ser1524) Antibody #9009 (上图) 和 BRCA1 Antibody #9010 (下图).

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat and K-562 cells using RAD50 Antibody #3427.Western blot 方法检测Jurkat和K-562细胞,使用的抗体为RAD50 Antibody #3427。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and K-562 cells using Mre11 Rabbit (31H4) mAb #4847.Western blot 方法检测HeLa和K-562细胞,使用的抗体为Mre11 Rabbit (31H4) mAb #4847。

Western Blotting

Western Blotting

Western blot analysis of extracts from wildtype M059K and DNA-PK deficient M059J cells using DNA-PK Antibody #4602.Western blot 方法检测野生型M059K和DNA-PK缺失的M059J细胞提取物,使用的抗体为DNA-PK Antibody #4602。

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or UV-treated (100 mJ, 4 hour recovery) 293 cells using Phospho-ATM (Ser1981) (D6H9) Rabbit mAb #5883 (upper) or ATM (D2E2) Rabbit mAb #2873 (lower).Western blot方法检测未处理或紫外(100 mJ, 恢复4 小时)处理的293细胞,使用的抗体为 Phospho-ATM (Ser1981) (D6H9) Rabbit mAb #5883 (上图) 或 ATM (D2E2) Rabbit mAb #2873 (下图).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or UV-treated (100 mJ, 4 hr recovery), using Phospho-ATM (Ser1981) (D6H9) Rabbit mAb (upper) or ATM (D2E2) Rabbit mAb #2873 (lower).

Background

Double strand DNA breaks (DSB) in mammalian cells can be repaired by the related mechanisms of non-homologous end-joining (NHEJ) and homologous recombination (HR). A DNA-dependent protein kinase composed of DNA-binding subunits Ku70 and Ku86 and the DNA-PKcs catalytic subunit mediates NHEJ repair. The Ku heterodimer binds free DNA ends and recruits DNA-PKcs to the break (1). DNA-PKcs signals areas of DNA damage and recruits additional proteins, such as the Artemis exo- and endonuclease that processes and primes the damaged sequence (2,3). Following replacement DNA synthesis, a ligase complex composed of DNA ligase IV and XRCC4 joins the repaired ends. XRCC4-like factor (XLF) is an essential ligase-associated repair factor that promotes gap-filling during NHEJ (4). Homologous recombination utilizes aligned homologous sequences as a repair template. The MRN complex, composed of Mre11, Rad50, and nibrin (p95/NBS1), plays a critical role in sensing, processing and repairing breaks (5). MRN interacts with BRCA1 and CtIP to facilitate 5’ resection of DSB DNA to generate 3’ ssDNA ends necessary for repair (6). DNA-binding protein Mre11 exhibits exonuclease and endonuclease activity and is largely responsible for ssDNA end processing (7). Interaction between the MRN complex and ATM kinase promotes association between the kinase and its substrates and likely leads to ATM activation (8). ATM acts a central controller of the cell cycle checkpoint by phosphorylating multiple targets, including c-Abl, BRCA1 and p95/NSB1. Activated c-Abl phosphorylates Rad52, which promotes Rad51 binding to ssDNA and subsequent annealing of ssDNA (7). 哺乳动物细胞中的双链DNA断裂(DSB)可以通过非同源末端连接(NHEJ)和同源重组(HR)进行修复。一种DNA依赖的蛋白激酶由DNA结合亚基Ku70和Ku86以及介导NHEJ修复的DNA-PKcs催化亚基组成。Ku异源二聚体结合自由的DNA末端和招募DNA-PKcs至断裂处(1)。 DNA-PKcs标志DNA损伤区域和招募额外的蛋白质如Artemis外切和内切核酸酶,处理和填补损坏的序列(2,3)。接下来替换DNA合成,DNA连接酶IV和XRCC4组成的连接酶复合物参与修复末端。XRCC4-样因子(XLF)是一个重要的连接酶相关修复因子,能够促进NHEJ期间的间隙填充(4)。同源重组利用均衡的同源序列作为修复模板。Mre11,Rad50和nibrin(p95/NBS1)组成的MRN复合物在传感、加工和断裂修复中起着至关重要的作用(5)。MRN与BRCA1和CtIP相互作用,以便切除5'DSB DNA以产生修复必需的3'单链DNA末端(6)。DNA结合蛋白Mre11具有外切酶和核酸内切酶活性,很大程度上负责单链DNA末端处理(7)。MRN复合物和ATM激酶之间的的相互作用能够促进激酶和其底物之间的关联并可能引起ATM活化(8)。ATM作为细胞周期检验点中央控制器的角色有赖于磷酸化多个靶标,包括c-Abl、BRCA1和p95/NSB1。活化的c-Abl能够磷酸化RAD52 Rad52,促进Rad51结合到单链DNA和随后单链DNA的退火(7)。

Application References

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

U.S. Patent No. 5,675,063.

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

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

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