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Rapid, Sensitive Detection of Proteins in Minigels With Fluorescent Dyes

Coomassie Fluor Orange, SYPRO® Orange, SYPRO Red, and SYPRO Tangerine Protein Gel Stains
  • Thomas H. Steinberg
  • Courtenay R. Hart
  • Wayne F. Patton
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

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.

Keywords

Sodium Dodecyl Sulfate Staining Solution Fluorescent Stain Single Vial Print Film 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2005

Authors and Affiliations

  • Thomas H. Steinberg
    • 1
  • Courtenay R. Hart
    • 1
  • Wayne F. Patton
    • 1
  1. 1.Molecular Probes, Inc.Eugene

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