Anti actin c 11 sc1615

Cells were lysed in ice-cold whole cell extract buffer containing 50 mM TrisHCl at pH 7.4, 250 mM NaCl, 0.1% Nonidet NP40, 5 mM EDTA and NaF 50 mM with a protease inhibitor cocktail (Sigma). Lysates were cleared by centrifuging at 12,000 rpm for 5 min. Cell lysates containing equal amounts of protein (30–70 µg) were resolved on 10–12% SDS-PAGE (polyacrylamide gel electrophoresis) gels. The proteins were then transferred to nitrocellulose membranes (PROTRAN, Schleicher and Shull). Immunoblotting was carried out with the following antibodies and visualized using Odissey FC Imaging System (Li-COR): anti-PLK1 (F-8) #sc17783, anti-actin (C-11) #sc1615 and anti-beta tubulin #sc9104 were provided by Santa Cruz Biotechnology. Anti-phospho-Histone H3 (Ser10) (6G3) #9706 was purchased from Cell Signaling Technology and anti-H2AX pSer139 #05-636 was purchased by Millipore. The horseradish peroxidase (HRP) conjugated secondary antibody anti-goat (sc-2354) was purchased from Santa Cruz Biotechnology (Heidelberg, Germany). The anti-rabbit (#1706515) and anti-mouse (#1706516) antibodies were purchased from BIO-RAD LABORATORIES S.r.l.