Anti cleaved parp asp214 rabbit monoclonal ab clone d64e10

24 h post-transfection, cell lysates were prepared using 1× cell lysis buffer (Cell Signaling) with 1 mM PMSF (Cell Signaling). 6–10 ug of protein lysate was run on a 4–20% SDS PAGE and transferred to Hybond-P or nitrocellulose membrane (GE Healthcare). The membrane was incubated for 20 min at room temperature (RT) in a 4% BSA (Omnipur) solution, washed, and incubated overnight (O/N) at 4 °C with one of the antibodies listed below. After washing and incubation for 1 h at RT with secondary antibody, the protein signal was detected using a Pierce ECL2 substrate (Thermoscientific) or Odyssey CLx imaging system (LI-COR Biosciences) according to the manufacturer’s instructions. Antibodies for immunoblotting were as follows: anti-PARP rabbit monoclonal Ab (clone 46D11; Cell Signaling), anti-cleaved PARP (Asp214) rabbit monoclonal Ab (clone D64E10; Cell Signaling), anti-GAPDH-peroxidase mouse Ab (Sigma), anti-RPS13 rabbit polyclonal Ab (Abnova), anti-RPS7, 10, 16, 19, 24, and 28, XRN2 Ab (Abcam), anti-RPS9, 14, and 15 Ab (Aviva), RPL7, 13a, 26, and XRN2 Ab (Abcam), anti-RPS6 mouse monoclonal Ab, anti-flag Ab (Cell signal).