Preparation method of highly-purified antibodies from antisera by using polyvinylidene difluoride membrane. Application to fatty acid binding protein in rat liver.

A simple method for preparation of highly-purified antibodies from antiserum was developed by using antigen-blotted polyvinylidene difluoride transfer membrane as an affinity matrix: The antigen was purified by SDS-polyacrylamide slabgel electrophoresis. Purified antibody preparations, which had been eluted from the Gly-HCl buffer (pH 2.5)-pretreated affinity matrix, by treatment with 1,4-dioxane, were more effective to obtain specifically stained band in immunoblot analysis than antiserum or immunoglobulin fractions purified by already reported methods. Antigen-blotted membranes were repeatedly available without remarkable release of antigen or decrease of antigenicity. This method was also applied to the examination of cross-reacting proteins with rat liver fatty acid binding protein in the cytosol fraction.