Abstract
A simple, rapid, and sensitive method for the fluorometric assay of proteins is described in this communication. Proteins are spotted on a Kieselguhr G plate and N-chlorinated by spraying with a solution of sodium hypochlorite. After removal of excess hypochlorite by aeration, the plate is sprayed with a solution of thiamine. The fluorescence of thiochrome, formed by the oxidation of thiamine with the N-chlorinated proteins, is then measured with a scanning fluorometer. Interfering substances including DNA are readily removed by the development of the plate with 2 m sodium chloride prior to the assay. The standard curves were linear in the range of 0.1 to 1.0 μg/spot of proteins, and the fluorescence intensity deviated only slightly among proteins.
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