Anti alpha tubulin sc 23948

Whole cells were lysed in a chilled lysis buffer (150 mM sodium chloride, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, and 50 mM Tris pH 8.0) with protease inhibitor cocktail (Roche, Switzerland). The protein concentration was determined using the Bradford assay. The same amount of protein was resolved by SDS-PAGE. After electrophoresis, the proteins were transferred onto nitrocellulose membranes (BioTrace NT Nitrocellulose membrane; PALL, USA). Then, the membrane was blocked for 1 h at room temperature using 5% skim milk in TBST buffer. The membrane was incubated overnight at 4°C with the following primary antibodies: anti-SETD5 (MBS8245298, 1:1,000; MyBioSource, USA), anti-alpha-tubulin (SC-23948, 1:10,000; Santa Cruz Biotechnology, USA), anti-PARP (9542, 1:1,000; Cell Signaling Technology, USA), anti-H3K27me3 (07-449, 1:1,000; Sigma-Aldrich), anti-H3K4me3 (39159, 1:1,000; Active Motif, Germany), anti-H3K9me2 (39375, 1:1,000; Active Motif), anti-H3K36me3 (ab9050, 1:1,000; Abcam, UK), anti-total H3 (ab1791, 1:1,000; Abcam), anti-phospho-STAT1 (9167, 1:1,000; Cell Signaling Technology), anti-STAT1 (9172, 1:1,000; Cell Signaling Technology), anti-MDA5 (5321, 1:1,000; Cell Signaling Technology), and anti-PKM (ab137791, 1:1,000; Abcam).