ROTI®Free Stripping Buffer, 1 l

The buffer offers a simple tool for removing antibodies and chemoluminescence staining from Western blot membranes, allowing the valuable membranes to be analysed several times. Excellent when several proteins are to be detected in the same extract, or e.g. when in addition to a specific analysis, proteins in the respective lanes have to be semi-quantified (Kaufmann et. al. (1987) Anal. Biochem. 161:89-95). Suitable for nitrocellulose and PVDF-membranes.

Wash the membrane in TBST-buffer (Art. No. 1061.1), incubate in ROTI^®Free Stripping-buffer in closed hood for 30 min at 56 °C (e. g. in shaking water bath). To strip AP-conjugates, we recommend incubation at 70 °C for 30-60 min. After washing the membrane twice in TBST for 20 min each time, it should first be blocked, before the first antibody is added. The stripping process can be checked by brief chemiluminescence staining after washing in TBST.
A slight reduction of the membrane-bound proteins and increase of the background may appear after repeated stripping. Store membranes after stripping wetpacked at +2 to +8 °C.