Improved Specificity of Western Blot Following Processing at 4°C Using BlotCycler

Western blot processed by BlotCyclerTM as compared to the traditional manual procedure. Western blots with cell extract from the clear cell renal carcinoma (ccRCC) cell line were incubated with the rabbit polyclonal anti‐PARP antibody (Cell Signal, Cat # 9542) using the traditional manual procedure (A) or BlotCyclerTM (B). The membranes were processed as follows: blocked with TBST‐5% dry milk for 1 hour, incubated with primary antibody overnight, washed 3X15 minutes with TBST, incubated with the anti‐rabbit secondary antibody (Jackson Immunochemicals, 1:4000 dilution) for 1 hour, and washed 3X15 minutes TBST. The bands were visualized using a chemiluminescence substrate.