Practice question

Stripping and reprobing constitutes essential procedure to reuse same membrane for detecting multiple different proteins. After initial detection, membrane is incubated in buffers containing SDS, beta-mercaptoethanol or dithiothreitol, and glycine at low pH or elevated temperature to dissociate and remove bound primary and secondary antibody complexes, while target proteins remain immobilized due to strong hydrophobic interactions with PVDF or nitrocellulose. Subsequent reprobing with antibody against different antigen, such as housekeeping protein beta-actin or phosphorylated isoform, allows normalization, conserves limited clinical or fractionated samples, and reduces inter-gel variability.