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A Rapid and Sensitive Method for the Quantitation of ...

ANALYTICAL BIOCHEMISTRY 72, 248-254 (1976). A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding MARION M. BRADFORD. Reproduction Research Laboratories, Department of Biochemistry, University of Georgia, Athens, Georgia 30602. Received September 11, 1975; accepted January 29, 1976. A protein determination Method which involves the binding of Coomassie Brilliant Blue G-250 to protein is described. The binding of the dye to protein causes a shift in the absorption maximum of the dye from 465 to 595 nm, and it is the increase in absorption at 595 nm which is monitored.

spectrophotometrically in a Bausch and Lomb Spectronic 200 uv spectrophotometer based on E~~,,+z = 6.6 (19,20), 5.3 (19,21), 20 (19,22) and 17.1 (23,24) respectively. Hemoglobin solutions were prepared gravimetrically. Preparation ofprotein reagent. Coomassie Brilliant Blue G-250 (100 mg)

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