Protease treatment of nitrocellulose-bound antigens enhances the sensitivity of western blots.
The sensitivity of Western blots is limited by the avidity of antibody-antigen interactions, and by problems of specificity in interactions between antibodies and antigens from different species. Using rat and human keratins as the antigens, and a set of antibodies against human and rat keratins, this study demonstrates that mild treatment of nitrocellulose blots with trypsin and/or pepsin enhances the sensitivity of the assay and permits the cross-species demonstration of antigens that are not otherwise detectable.
Pan
J
J
Department of Anatomy, University of British Columbia, Vancouver, Canada.
Auersperg
N
N
D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D002478:Cells, Cultured; D003101:Collodion; D010450:Endopeptidases; D005260:Female; D006801:Humans; D007633:Keratins / Q000032:analysis; D010434:Pepsin A; D051381:Rats; D012680:Sensitivity and Specificity; D014357:Trypsin
D007633:Keratins; D003101:Collodion; D010450:Endopeptidases; D014357:Trypsin; D010434:Pepsin A