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Estimation of molecular weight by polyacrylamide gel electrophoresis using heat stable fluorophors

✍ Scribed by Bruce O. Barger; Fred C. White; Judith L. Pace; David L. Kemper; W.L. Ragland

Publisher: Elsevier Science
Year: 1976
Tongue: English
Weight: 464 KB
Volume: 70
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(76)90453-x

No coin nor oath required. For personal study only.

✦ Synopsis

The utilization of 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF) as a fluorescent label for proteins in SDS polyacrylamide gel electrophresis is described. The procedure is very sensitive and detects disks containing as little as 1 ng of protein. The fluorescence signal is linear with concentration of protein to 500 ng. Emphasis is placed on the use of MDPF-labeled proteins as standards for the estimation of molecular weights of fluorescamine-labeled proteins. Data are provided which demonstrate the need for only three standard proteins in the molecular weight curve. Finally, the relative mobilities of proteins from nonreplicate determinations exhibited an average variation of only 3.9%.

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