SYPRO Orange Fluorescent Staining of Protein Gels
Adapted from Proteins and Proteomics by Richard J. Simpson. CSHL Press, Cold Spring Harbor, NY, USA, 2003.INTRODUCTION
This method for staining protein gels uses a fluorescent dye, SYPRO Orange, which is very sensitive to protein (4-10 ng protein/band). SYPRO Orange cannot be visualized with the naked eye and thus a fluorescence scanner is required for detection of protein bands (the excitation maxima [Ex] are 300/472 nm and the emission maximum [Em] is 570 nm). SYPRO Red fluorescent protein stain uses the same binding principle as SYPRO Orange; hence, this protocol can also be used for SYPRO Red stain (Ex are 300/550 nm and Em is 630 nm). This staining method is mass-spectrometer-compatible.
