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Quantitative assay for submicrogram amounts of protein

โœ Scribed by Melvin L. Goldberg

Publisher
Elsevier Science
Year
1973
Tongue
English
Weight
426 KB
Volume
51
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

An ultrasensitive

protein assay, linear between 0.01 and 0.2 pg protein, is described. In this assay, copper is complexed to protein, excess copper is removed by adsorption to a small Sephadex column, and the copper-protein complex is destroyed by digestion with hydroperoxide.

Phenol and chloramine-T are added to the reaction mixture and the copper catalyzes the production of a color-producing reaction between the two compounds. The assay is not affected by low levels of phosphate, Tris, metals, or a tenfold excess of nucleic acid over protein. Reducing agents, sucrose, certain anions, high salt concentrations, and EDTA seriously interfere. The method is about 500 times as sensitive as that of Lowry et al. [

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