Rabbit anti phospho s6 ribosomal protein ser235 236 antibody

Whole cell lysates were isolated using standard RIPA buffer containing proteases and phosphatases and quantified using the Lowery protein assay (BioRad). 50 μg of lysates from BCPAP cells treated with DMSO vehicle or increasing concentrations (25 nM, 50 nM, or 100 nM) of TM for 7 days or TM (EC_6.25, 25 μM), vemurafenib (EC_6.25, 6.25 μM) or both drugs at the same concentrations for 7 days were resolved by SDS-PAGE and immunoblotted with a rabbit anti-phospho(Thr 202/Tyr 204)-ERK1/2 antibody (Cell Signaling Technology, antibody # 3700 at a 1:1000 dilution), a mouse anti-ERK1/2 antibody (Cell Signaling Technology, antibody # 9101 at a 1:1000 dilution), a mouse anti-HA-Tag antibody (Cell Signaling Technology, antibody # 2367 at a 1:1000 dilution), a rabbit anti-phospho-S6 ribosomal protein (Ser235/236) antibody (Cell Signaling Technology, antibody #4858S at a 1:1000 dilution), a rabbit anti-S6 ribosomal protein (Cell Signaling Technology, antibody #2217 at a 1:1000 dilution), or a mouse anti-β-tubulin (Sigma-Aldrich, antibody # 2367 at a 1:5000 dilution) followed by a goat anti-rabbit IgG (Cell Signaling Technology, antibody # 7076) or a goat anti-mouse IgG (Cell Signaling Technology, antibody # 7074) horseradish peroxidase-conjugated secondary antibody and visualized using enhanced chemiluminescence detection (Cell Signaling Technology).