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Quantitation of collagen in polyacrylamide gels by fluorescent scanning of MDPF1-labeled proteins: An improvement over densitometric scanning of gels stained by coomassie blue

1978; Elsevier BV; Volume: 90; Issue: 1 Linguagem: Inglês

10.1016/0003-2697(78)90009-x

1096-0309

Ronald Goldberg, George C. Fuller,

Osteoarthritis Treatment and Mechanisms

Purified α chains of collagen were made fluorescent by coupling with 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF) and then electrophoresed on sodium dodecyl sulfate-polyacrylamide gels. The migration of MDPF-labeled collagen was similar to unlabeled collagen α chains except that MDPF-α1(I) migrated closer to MDPF-α2. The area under the peaks recorded from fluorometric scanning of MDPF-labeled α1(I), α2, and α1(III) was linear from 10−5 to 10−8 g. The standard curves for the three α chains were similar. The results from nonreplicate determinations had ±6% SE. This method is an improvement over staining with Coomassie blue. It has a greater sensitivity, peak area is independent of migration distance, has a wider range of linearity, and permits observation of bands during electrophoresis with quantitation immediately after electrophoresis.