Chloramine T-mediated radiolodination of proteins on intact polyacrylamide slab gels

Abstract

A simple, inexpensive method is described here for the radioiodination of proteins on intact, developed polyacrylamide gels. The method is based on the chloramine T iodination procedure which yields proteins containing 125I label specifically in tyrosine residues. When employed with intact polyacrylamide gels, our method allows detection of proteins in amounts too small to be observed by chemical stains, including Coomassie blue. The procedure should, therefore, be useful for analyzing protein mixtures where only a small amount of material is available or for assaying trace contaminants in purified protein preparations. Proteins radioactively labeled by our method are suitable for further analysis by proteolytic cleavage followed by peptide mapping or “fingerprinting.”