Multiple immunoblot: A sensitive technique to stain proteins and detect multiple antigens on a single two‐dimensional replica

Walter Steffen, Richard W Linck

Research output: Contribution to journalArticle

13 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)714-718
Number of pages5
JournalELECTROPHORESIS
Volume10
Issue number10
DOIs
StatePublished - 1989

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