Immunofluorescent protein detection in Western blotting

This report describes the detailed procedures for Western blot analysis using fluorescent antibodies. After electrophoresis and subsequent electroblotting, the fluorescentlabeled antibodies were visible upon ultraviolet illumination of the polyvinylidene fluoride (PVDF) membranes and could then be photographed to give an accurate record of the blots. Fluorescent labeling allows for photographic records of the blot without the need for subsequent staining steps. In an effort to evaluate this method in the study of immunogenic antigens, this study describes a study on BLV (Bovine Leukemia Virus) tumor antigens. The fluorescentlabeling of antibodies for detection by immunoblotting was found to be a very sensitive and reliable alternative to the conventional methods ofWestern blots.

Keywords:

immunofluorescence , immunoblotting , Western blotting , FITC , PVDF