α tubulin rabbit mab

Total protein was collected by lysing adherent cells with RIPA buffer (Sigma, cat. #R0278) supplemented with phosphatase and protease inhibitors (Thermo Scientific, cat. #78440). Protein concentration was measured using the Bradford reagent (Thermo Scientific, cat. #23236). An equal amount of protein was loaded for immunoblotting. After electrophoresis, the protein was transferred to the PVDF transfer membrane (Merck, cat. #0000240973), followed by a 5% BSA blocking buffer, and incubated overnight with primary antibodies. After washing with TBST (TBS with .1% Tween), anti‐rabbit second antibodies (Cell Signaling Technology, cat. #7074) conjugated with HRP‐conjugated were used to probe the membranes. Samples were developed by Chemiluminescent Substrate (Thermo Scientific, cat. #34577) and exposed by Tanon 5200 Multi‐system (Tanon Co.). Densitometry was performed using ImageJ software (National Institutes of Health), and samples were normalized to internal controls. Primary antibodies used in this study are CDK12 antibody (Cell Signaling Technology, cat. #11973), Phospho‐Rpb1 CTD (Ser2/Ser5) Rabbit mAb (Cell Signaling Technology, cat. #13546), β‐Actin Rabbit mAb (Cell Signaling Technology, cat. #8457), GAPDH Rabbit mAb (Cell Signaling Technology, cat. #2118), α‐Tubulin Rabbit mAb (Cell Signaling Technology, cat. #2125) and recombinant Anti‐FACL4 antibody (Abcam, cat. #EPR8640).