BOVINE SERUM-ALBUMIN (BSA) AS A REAGENT AGAINST NONSPECIFIC IMMUNOGOLD LABELING ON LR-WHITE AND EPOXY-RESIN

The purpose of this study was to examine how different incubation time s with different concentrations of bovine serum albumin (BSA) affect t he amount of non-specific immunogold labeling on epoxy sections and LR -White sections. Immunogold labeling was performed on epoxy sections a nd LR-White sections of renal tissue with IgG-deposits and fibrin clot s, and the antibodies used were anti-IgG and anti-fibrinogen, respecti vely. The sections were incubated with different concentrations of BSA prior to application of primary antibodies, and the length of this pr e-incubation step varied between 0 and 4 h. During the incubation with primary antibodies, BSA was added in the same concentration as in the pre-incubation step. The results showed that the non-specific labelin g on the resin decreased significantly when the concentration of BSA o r the length of the preincubation step was increased. The non-specific labeling was usually higher on the epoxy resin than on the LR-White r esin when using the same conditions with respect to BSA. But, when the preincubation step with BSA lasted 4 h, the non-specific labeling was somewhat lower on epoxy resin than on the acrylic LR-White resin, wit hout respect to the concentration of BSA. The specific labeling for bo th fibrinogen and IgG decreased slightly when the concentration of BSA and incubation time increased, probably due to the steric hindrance p erformed by BSA molecules on the section. Blocking procedures with at least 1 h incubation time for the blocking step with at least 5% BSA a re recommended for both epoxy and LR-White sections. (C) 1997 Elsevier Science Ltd.