Journal
ELECTROPHORESIS
Volume 25, Issue 15, Pages 2506-2510Publisher
WILEY
DOI: 10.1002/elps.200406004
Keywords
fluorescent stain; protein detection; proteomics; SYPRO ruby
Funding
- NCI NIH HHS [R33 CA093292] Funding Source: Medline
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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.
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