Multiple immunoblot: a sensitive technique to stain proteins and detect multiple antigens on a single two-dimensional replica |
| |
Authors: | W Steffen R W Linck |
| |
Institution: | Department of Cell Biology and Neuroanatomy, University of Minnesota, Minneapolis 55455. |
| |
Abstract: | A multiple immunoblotting technique was developed to positively identify up to three different antigens on a single nitrocellulose replica of a two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel. Three highly sensitive immunoblot assays were selected, including: horseradish peroxidase/luminescence, alkaline phosphatase, and silver-enhanced immunogold. As a major advantage, the method permits a simultaneous detection of up to three different antigens without eluting the antibody-dye complex between staining of single polypeptides, thus providing a highly accurate identification of closely migrating components. The staining procedure is summarized in a flow chart. In addition to the multiple immunoblot staining, some suggestions are provided for a sensitive protein staining. |
| |
Keywords: | |
|
|