Anti rad51 gtx100469

The primary antibodies used in this study were as follows: anti-RECQL4 antibody was prepared as previously described (24 ). Other antibodies used in this study included anti-pATM (Ser-1981, #4526, Cell Signaling), anti-MRE11 (GTX30294, GeneTex), anti-NBS1 (A7703, ABclonal), anti-RAD51 (GTX100469, GeneTex), anti-USP2 (A10399, ABclonal), anti-USP28 (A9292, ABclonal), anti-γH2AX (A300-081A, Bethyl and 05-636, EMD Millipore), anti-Lamin B1 (ab16048, Abcam), anti-GFP (sc-9996, Santa Cruz), anti-FLAG M2 (Sigma), and anti-HA (AE008, ABclonal). Alexa Fluor 488 antimouse IgG (A11001, ThermoFisher), Alexa Fluor 488 anti-rabbit IgG (A11008, ThermoFisher), Alexa Fluor 594 antimouse IgG (A11005, ThermoFisher), and Alexa Fluor 594 anti-rabbit IgG (A11012, ThermoFisher) were used as secondary antibodies for immunofluorescence microscopy.

GFP-CtIP, full-length and truncated FLAG–CtIP were generously provided by Dr. Junjie Chen (MD Anderson Cancer Center, TX). FLAG–ASF1A was generously provided by Dr. Zhiguo Zhang (Columbia University, NY). Full-length and truncated FLAG–USP52 and Myc-USP52 were generously provided by Dr. Lei Shi (Tianjin Medical University, Tianjin, China). FLAG–USP52 S1003A and S469A, FLAG–CtIP K62R, K760R, K782R, and 2KR mutants were generated by site-directed mutagenesis (Stratagene). Flag–CtIP–T847E was purchased from Addgene. CHX, IgG agarose, anti-FLAG agarose, 3×FLAG peptide and ATM inhibitor KU55933 were purchased from Sigma Aldrich. Olaparib was purchased from Toronto Research Chemicals. Anti-USP52 (ab241505, 1:1000) was purchased from abcam; anti-CtIP (Thr847) (p1012-847, 1:2000), and anti-CtIP (Thr327) (p1012-327, 1:2000) were purchased from PhosphoSolutions; anti-RPA32 (sc-56770, 1:2000), anti-Ub (sc-8017, 1:2000) and anti-CtIP (sc-271339, 1:1000 for WB) were purchased from Santa Cruz; anti-FLAG (F1804, 1:2000) was purchased from Sigma; anti-pS345 Chk1 (2348, 1:1000), anti-Myc Tag (2276, 1:2000) and anti-SQ/TQ motif (9607, 1:1000) were purchased from CST; anti-CtIP (61141, 1:1000 for IF) was purchased from Active Motif; anti-GAPDH (60004-1-lg, 1:2000) was purchased from Proteintech; anti-Rad51 (GTX100469, 1:1000) was purchased from Genetex.

Antibodies used in this study were as follows: anti-USP15 (A300-923A, WB: 1:1000), anti-BARD1 (A300-263A, WB: 1:1000; IF: 1:200), and anti-RAP80 (A300-764A, WB: 1:1000; IF: 1:100) were purchased from Bethyl Laboratory. Anti-HP1γ (MABE656, WB: 1:2000). Mouse anti-γ-H2AX (05-636, IF: 1:500), mouse anti-MDC1 (05-1572, IF: 1:300) and mouse anti-FK2 (04-263, IF: 1:200) were purchased from Millipore. Anti-RPA 70 (#2267, IF: 1:200), rabbit anti-γ-H2AX (#9718, IF: 1:400), and rabbit anti-HA (#3724, WB: 1:2500; IF: 1:300) were from Cell signaling Technology. Anti-Rad51 (GTX100469, IF: 1:200) were from Genetex. Anti-BRCA1 (D9, IF: 1:50) and anti-His tag (sc8036, WB: 1:1000) were from Santa Cruz. Anti-poly PAR (4336-BPC-100, WB: 1:1000) were from Trevigon. Anti-53BP1 (NB100-304, WB: 1:1000; IF: 1:300) were from Novus Biologicals. Anti-RNF8 (14112-1-AP, IF: 1:100) were from Proteintech. Rabbit anti-FLAG (F7425, WB: 1:2500) and mouse anti-FLAG (F3165, WB: 1:2500) were from Sigma-Aldrich. Mouse anti-HA (901501, WB: 1:2500; IF: 1:100) were from BioLegend.