Technical Support FAQ


Technical Support FAQ


Can membranes be stripped after using the Immun-Star HRP chemiluminescent kits? What is the recipe for the stripping buffer?


Membranes can be stripped and reused if kept wet after starting the western blot process. There are several protocols for stripping the membrane for a second probing with a different antibody. One of the most common calls for the use of high temperatures (70ºC) with 2% SDS and 100 mM beta-mercaptoethanol (BME) in 62.5 mM Tris-HCl, pH 6.7, for 30 min to destroy the antigen-antibody complex. Other methods use a very high (9.5) or very low (2.2) pH with the presence of detergents such as Tween 20 or SDS.

A harsher method, which will desorb proteins from polyvinylidene difluoride (PVDF) or nitrocellulose membranes, is outlined in an article by Montelaro (Protein antigen purification by preparative protein blotting, Electrophoresis 8, 432-438, 1987). In this method, 40% acetonitrile or pyridine is used to desorb protein from PVDF.

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