Rapid, Sensitive Detection of Proteins in Minigels With Fluorescent Dyes
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The advent of polyacrylamide gel electrophoresis (PAGE) in the minigel format (i.e., gel dimensions in the range of 6 × 9 cm or 8 × 8 cm × 0.75 to 1.5 mm thick), the widespread use of precast minigels, and the commercialization of colloidal Coomassie® Blue G-250 staining formulations have raised expectations for rapid, sensitive postelectrophoresis staining methods. The proprietary commercially available fluorescent stains Coomassie Fluor− Orange protein gel stain, SYPRO® Orange dye, SYPRO Red dye, and SYPRO Tangerine dye (Molecular Probes, Inc., Eugene, OR) offer sensitivity and ease of use that rival or surpass colloidal colorimetric staining. These stains are characterized by high sensitivity, a 3-log linear dynamic range, and a simple staining protocol, allowing rapid, accurate quantitation of proteins in gels. Staining is reversible and can be followed by staining with Pro-Q® Diamond phosphoprotein stain or Pro-Q Emerald glycoprotein stain for detection of posttranslational modifications. Staining can also be followed by detection with other total protein stains, including SYPRO Ruby stain or colorimetric stains such as Coomassie Blue or silver. A common feature of these fluorescent protein gel stains is a bimodal excitation spectrum, with maxima in the ultraviolet (UV) and visible ranges, resulting in versatility of use on a broad range of imaging platforms.
KeywordsSodium Dodecyl Sulfate Staining Solution Fluorescent Stain Single Vial Print Film