Stripping buffer for removal of high-affinity antibodies, as well as chemoluminescence staining, from Western blot membranes. Due to the low pH of the buffer, the antibody conformation is altered, therefore gently removing all molecules bound to the epitopes (Legocki et. al. (1981) Analyt. Biochem. 111:385-92). Specially designed additives enhance the efficiency in removing those antibodies requiring long stripping times or high incubation temperatures with standard stripping buffer systems. Advanced formulation without β-mercaptoethanol or DTT for use at room temperature or heated. Suitable for nitrocellulose and PVDF membranes.

Following rinsing steps in PBST- or TBST buffer, the membrane is incubated in ROTI^®Free Stripping-Puffer 2.2 plus for 30-60 mins. at room temperature. In case of very persistent antibodies incubation in heated buffer is allowed. After washing it again, the membrane may be blocked and probed anew. The particular time needed for complete stripping depends on the respective antibody/epitope pairing.