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).
Western blot analysis of extracts of HeLa, NCCIT and PYS2 cells using ATM (D2E2) Rabbit mAb.
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 blot analysis of extracts from M059K (DNA-PK wildtype) and M059J (DNA-PK deficient) cells, using DNA-PK Antibody.
Confocal immunofluorescent analysis of HeLa cells using Ku80 (C48E7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
Immunohistochemical analysis of frozen SKOV-3 xenograft using Mre11 (31H4) Rabbit mAb.
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 blot analysis of extracts from Jurkat and K562 cells, using RAD50 Antibody.
Western blot analysis of extracts of various cell lines using XLF Antibody.
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.
Immunohistochemical analysis of frozen SKOV-3 xenograft using Ku80 (C48E7) Rabbit mAb.
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).
Immunohistochemical analysis of paraffin-embedded human glioblastoma using Ku80 (C48E7) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded lung carcinoma, using Mre11 (31H4) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Ku80 (C48E7) Rabbit mAb.
Western blot analysis of extracts from HeLa and K562 cells, using Mre11 Rabbit (31H4) mAb.
Immunohistochemical analysis of paraffin-embedded human melanoma using Ku80 (C48E7) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human GIST using Ku80 (C48E7) Rabbit mAb.
Western blot analysis of cell extracts from HeLa, A549 and COS cells using Ku80 (C48E7) Rabbit mAb.
|Phospho-ATM (Ser1981) (D6H9) Rabbit mAb 5883||20 µl||
|ATM (D2E2) Rabbit mAb 2873||20 µl||
||H M||350||Rabbit IgG|
|Phospho-BRCA1 (Ser1524) Antibody 9009||20 µl||
|DNA-PKcs Antibody 4602||20 µl||
|Ku80 (C48E7) Rabbit mAb 2180||20 µl||
||H Mk||86||Rabbit IgG|
|Mre11 (31H4) Rabbit mAb 4847||20 µl||
|Phospho-p95/NBS1 (Ser343) Antibody 3001||20 µl||
||H Mi M||95||Rabbit|
|Rad50 Antibody 3427||20 µl||
|XLF Antibody 2854||20 µl||
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
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 two western blots with each antibody.
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.
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. ATM (D2E2) Rabbit mAb is produced by immunizing animals with recombinant 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, 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.
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).
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