Superlumia ecl hrp substrate kit

Total cellular protein was extracted using RIPA lysis buffer containing protease inhibitor cocktail (Beyotime, China). The extracted proteins were separated by SDS-PAGE and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, NY), which were subsequently blocked with a 5% solution of nonfat milk for 1 h. The membranes were then incubated with primary antibodies at 4°C overnight, followed by secondary antibody incubation for 1 h at room temperature. The proteins were visualized with a SuperLumia ECL HRP Substrate Kit (Millipore, NY) and detected using an LAS4000 chemiluminescence detection system (Fuji, Tokyo, Japan). The primary antibodies used were as follows: anti-β-actin (1:1,000, AF5001, Beyotime), SREBP1 (1:1,000, 14088-1-AP, Proteintech), and SCD1 (1:500, sc-515844, Santa Cruz Biotechnology). The anti-β-actin antibody was used as an internal control.

Total cellular protein was extracted using RIPA lysis buffer containing protease inhibitor cocktail (Beyotin, China). The extracted proteins were separated by SDS-PAGE and transferred onto PVDF membranes (Millipore, NY, USA), which were subsequently blocked with a 5% solution of skim milk for 1 h. The membranes were then incubated with primary antibodies at 4 °C overnight, followed by secondary antibody incubation for 1 h at room temperature. The proteins were visualized with a SuperLumia ECL HRP Substrate Kit (Millipore, NY, USA) and detected using an LAS4000 chemiluminescence detection system (Fuji, Tokyo, Japan). The primary antibodies used were as follows: anti-β-actin (1:1000, Beyotin, AA128) and anti-E2F8 (1: 500, sc-514064, Santa-Cruz Technology). An anti-β-actin antibody was used as an internal control.