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Staining of phospho-proteins on acrylamide gel electropherograms
Authors:J A Cutting  T F Roth
Institution:1. Department of Biology, University of California, San Diego, La Jolla, California, USA;2. Department of Biological Sciences, University of Maryland, Baltimore County, Catonsville, Maryland 21228 USA
Abstract:The paper describes a method for staining the phosphate moiety of phospho-proteins after electrophoresis on polyacrylamide gels. The method depends on hydrolysis of the phospho-protein phosphoester linkage using dilute base in the presence of calcium ions. The gel containing the newly formed insoluble calcium phosphate is then treated with ammonium molybdate in dilute nitric acid. The resultant insoluble nitrophospho-molybdate complex is finally stained with the basic dye, methyl green, dissolved in dilute acetic acid. After destaining, a green band is observed on the gel at the locus of the phospho-protein. One nanomole of protein-bound phosphate is detectable.
Keywords:To whom all correspondence and reprint requests should be addressed  
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