Anti p53 antibody do 1 sc 126

Whole cell lysate was prepared in RIPA lysis buffer with 1X Laemmli buffer, then denatured by heating at 100°C for 5 minutes. Denatured proteins were separated in SDS-PAGE gel and transferred onto PVDF membrane. The PVDF membrane was incubated in TBST with 5% nonfat dry milk for one hour at room temperature prior to incubating with primary antibody of interest overnight at 4°C with gentle rotation. Appropriate horse radish peroxidase conjugated secondary antibody was applied for one hour at room temperature. Signal was developed by applying SuperSignal West Pico chemiluminescence substrate and captured using the ChemiDoc imaging instrument. Primary antibodies used in this study: anti-AHR antibody (BML-SA210 – Enzo Biochem, Farmingdale, NY), anti-p27Kip1 antibody (610242 – BD Biosciences), anti-p53 antibody (DO-1 sc-126 – Santa Cruz Biotechnology, Dallas, TX), anti-p21 antibody (2947S – Cell Signaling Technology, Danvers, MA), and anti-GAPDH antibody (sc-36502, Santa Cruz).