Atr sc 1887

Collected cells were suspended in SDS sample buffer, boiled for 5 min, separated by SDS-PAGE and transferred to a polyvinylidene difluoride membrane. Membranes were incubated overnight with primary antibodies, followed by 1 h incubation with secondary antibodies. The antibodies used for western blotting were ATR (sc-1887; Santa Cruz Biotechnology), phospho-ATR Thr1989 (128145; GeneTex, Inc.), Chk1 (C9358; Sigma-Aldrich), phospho-Chk1 Ser345 (2348; Cell Signaling Technology), Cdk1 (sc-54; Santa Cruz Biotechnology), phospho-Cdk1 Tyr15 (9111; Cell Signaling Technology), Cdc25C (sc-13138; Santa Cruz Biotechnology), phospho-Cdc25C Ser216 (9528; Cell Signaling Technology), Cleaved-caspase3 (9661; Cell Signaling Technology), γH2AX (61796; GeneTex, Inc.) and β-actin (ab6276; Abcam).

Antibody to Phospho‐S345‐Chk1 (#2348) and Phospho‐S10‐Histone H3 (#53348) was purchased from Cell Signaling Technology (Danvers, MA, USA); BRCA1 (sc‐6954), Cyclin A (sc‐2712682), Chk1 (sc‐8408), and ATR (sc‐1887) from Santa Cruz (Dallas, TX, USA); p53 (Ab‐6, Clone DO‐1, MS‐187‐P0) from Thermo Fisher Scientific; 53BP1 (MAB3802) and γH2AX S139 (05‐636) from MilliporeSigma (Burlington, MA, USA); and FANCD2 (NB100‐182) from Novus Biologicals (Centennial, CO, USA). The polyclonal anti‐Timeless antibody was generated previously [28 (link)].