Antibody-based Analysis of DNA Damage Repair

Antibodies against the following proteins were used for immunofluorescence (IF) and Western blotting (WB): γH2AX (05–636 JBW301; Upstate Biotechnology), CENPF (ab5; Abcam), RPA (NA-18, Ab-2; Calbiochem for IF, LS-C38952; LifeSpan BioSciences for WB), pRPA S4/8 (A300-245A; Bethyl Laboratories), p DNA-PKcs S2056 (ab18192), DNA-PKcs (12311; Cell Signaling), Ku80 (2180 (C48E7); Cell Signaling), pKAP-1 S824 (A300-767A; Bethyl Laboratories), KAP-1 (ab3831; Abcam), MRE11 (12D7, GTX70212; GeneTex), NBS1 (PC269, Ab-1; Oncogene), RAD50 (3427; Cell Signaling), pATM S1981 (2152–1; Epitomics), ATM (GTX70103 2C1; GeneTex), CtIP (A300-488A; Bethyl Laboratories), and β-tubulin (ab21058; Abcam). Ku70, XLF, XRCC4, and Ligase IV antibodies were kind gifts from Dr. Penny Jeggo. Immunoblotting was performed as described previously [17 (link)].

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Nakajima N.I., Hagiwara Y., Oike T., Okayasu R., Murakami T., Nakano T, & Shibata A. (2015). Pre-Exposure to Ionizing Radiation Stimulates DNA Double Strand Break End Resection, Promoting the Use of Homologous Recombination Repair. PLoS ONE, 10(3), e0122582.

Publication 2015

CENPF pDNA-PKcs S2056 DNA-PKcs pKAP-1 S824 RAD50 pATM S1981

Antibodies Cenpf Ctip Dna pkcs Gifts Immunofluorescence Iv antibodies Kap 1 Ku70 Ligase Oncogene Pkcs Proteins Rad50 Tubulin Xrcc4

Corresponding Organization :

Other organizations : The Open University of Japan, Gunma University

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Variable analysis

independent variables

Antibodies against the following proteins: γH2AX, CENPF, RPA, pRPA S4/8, pDNA-PKcs S2056, DNA-PKcs, Ku80, pKAP-1 S824, KAP-1, MRE11, NBS1, RAD50, pATM S1981, ATM, CtIP

dependent variables

Immunofluorescence (IF) and Western blotting (WB) results

control variables

Immunoblotting procedure (performed as described previously [17])
Ku70, XLF, XRCC4, and Ligase IV antibodies (kind gifts from Dr. Penny Jeggo)

controls

Positive controls: Not explicitly mentioned.
Negative controls: Not explicitly mentioned.

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