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Optimized conditions for diluting and reusing a fluorescent protein gel stain
Author(s) -
Ahnert Nancy,
Patton Wayne F.,
Schulenberg Birte
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200406004
Subject(s) - fluorescence , stain , chromatography , chemistry , fluorescent labelling , reuse , staining , biology , ecology , genetics , quantum mechanics , physics
This study elucidates the optimum conditions at the minimum cost for using SYPRO Ruby protein gel stain. It deals with the effects of gel fixation and staining times, as well as dilution and reuse of SYPRO Ruby protein gel stain in one‐dimensional (1‐D) gels. Signal strength and dynamic range were highest in gels that were fixed thoroughly before staining, followed by overnight staining. Using the optimized protocol, dilution or reuse of the stain reduces the dynamic range and signal intensity. Sensitivity remains high if the stain is reused up to two times, but signal intensity is reduced up to 2.5‐fold in twice used stain. Sensitivity also remains high if the stain is diluted 1:2 in water, but signal intensity is reduced up to 6‐fold. Of the two options, reuse or dilution, reuse better retains signal intensity and dynamic range.