Anti rank antibody

The cells were harvested and lysed using ice-cold lysis buffer (Beyotime, Shanghai, China). The protein concentration was then determined using a protein assay kit (Keygentec, Nanjing, China). The denatured proteins (20 μg) were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). After blocking, the membranes were incubated at 4°C overnight with anti-TAZ antibody, anti-RANKL antibody, anti-RANK antibody, and anti-GAPDH antibody (Abcam, Cambridge, MA, USA). Next, the membranes were incubated with secondary antibodies for 2 h at 25°C. The bound proteins were visualized by enhanced chemiluminescence (Thermo Scientific) and detected using an imaging system (DNR Bio-Imaging Systems Ltd., Jerusalem, Israel), with GAPDH as the loading control.