Developing a dye-based method of protein quantitation on nitrocellulose

Abstract

time during the incubation periods for discussion of the blocking, washing, first antibody, conjugate, and detection steps. Moreover, this exercise allows for inclusion of the SDS-PAGE and transfer steps as separate laboratory exercises preceding the immunodetection phase. If protein gel electrophoresis and transfer equipment are not available, hybridoma supernatant can be directly added to nitrocellulose membrane strips using a dot-blot apparatus, with each protein-dot representing a different simulated target protein. This laboratory exercise is an effective procedure for demonstrating the theory and practice of the Western blot used in serodiagnosis.