Bicinchonininc acid protein assay kit

Total proteins were extracted from the xenograft tumors of different groups by ice-cold radio immunoprecipitation assay lysis buffer (Beyotime, Shanghai, China). After centrifuged at 12,000 rpm for 30 min, the lysate supernatants were harvested. The total protein levels were measured by the Bicinchonininc Acid Protein Assay Kit (Beyotime, Shanghai, China) according to the manufacturer's instructions.

After sample collection, total proteins were extracted and quantified using bicinchonininc acid protein assay kit (Beyotime, P0009, Shanghai, China). The equal proteins were resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to NC membrane. The membrane was blocked by 3% BSA contained in Tris Buffered saline Tween (TBST, 20 mM Tris–HCl, pH 8.0; 150 mM NaCl; 0.05% Tween-20) buffer for 2 h at room temperature, followed by incubation with corresponding primary antibody at 4 °C overnight. Then, the membranes were incubated with corresponding secondary antibodies for 1 h. Finally, the protein bands were imaged using an enhanced chemiluminescence system (ECL; Millipore, P90720, massachusetts, USA). Image J was applied to calculate protein levels.