Lifr sc 659

Total protein was extracted from the HCC cell lines with ice-cold radioimmunoprecipitation-assay lysis buffer, then separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were transferred to polyvinylidene difluoride (PVDF) membranes which were blocked in 5% non-fat powdered milk for 2h. Next, the membranes were incubated in primary detection antibodies overnight at 4°C and subsequently incubated with secondary detection antibodies for 2h at room temperature. The following primary antibodies were used to incubate membranes: LIFR (sc-659, Santa Cruz Biotechnology); GAPDH (#ab181602, Abcam). Protein expression levels were visualized using an enhanced chemiluminescence (ECL) detection system after washing the membranes three times with Tris-buffered saline-Tween (TBS-T) buffer.