Multiplexed Proteomics Technology
Invitrogen's Molecular Probes business has developed a suite of compatible technologies that provides a more complete picture of the proteome than has ever before been possible. Three technologies — SYPRO® Ruby protein gel stain, Pro-Q® Diamond phosphoprotein gel stain and the Pro-Q® Emerald glycoprotein stains — can be used individually or in series to detect total protein, phosphoproteins, and glycoproteins, respectively, using a single protein sample separated by 2D gel electrophoresis. Unambiguous spot matching of phosphoproteins and glycoproteins is made simple by direct comparison with the total protein profile stained with SYPRO® Ruby stain.
Proteins from either normal liver cells (left) or liver tumor cells (right) were separated using 2D gel electrophoresis and stained with Pro-Q® Diamond phosphoprotein gel stain (blue), Pro-Q® Emerald 300 glycoprotein stain (green) and then SYPRO® Ruby protein gel stain (red). The gel was imaged after each staining and the digital images were overlaid.
Multiplexed Proteomics Technology Applied to 1D Gels is a Time- and Labor-Saving Prescreening Process
Almost all modern proteomics studies involve some type of prefractionation sample analysis. There are often multiple sample fractions to analyze, and it is usually not clear which fractions will have the most interesting proteins. We recommend running all sample fractions on simple 1D gels (10 to 15 fractions per gel), and then staining the 1D gel for phosphoproteins, glycoproteins, and total proteins using the Pro-Q® Diamond, Pro-Q® Emerald, and SYPRO® Ruby protein gel stains, respectively. In this manner, it is possible to quickly determine the presence of phosphorylated and glycosylated proteins in a fraction, in relation to total-protein content. We have recently completed a study of this type and succeeded in rapidly identifying novel phosphorylated proteins within mitochondria (view an abstract or download the accepted manuscript at JBC Online) by prescreening multiple fractions from a sucrose density gradient using Pro-Q® Diamond and SYPRO® Ruby protein gel stains.
Used on its own, each of these state-of-the-art detection technologies brings you the following benefits:
- Simple, streamlined staining method
- High sensitivity
- Broad linear quantitation range
- Compatibility with many instruments, including gel scanners and mass spectrometers
Used together, the Multiplexed Proteomics technologies streamline the process of rapid, large-scale data acquisition and make possible, for the first time, a comprehensive look at the proteome, the phosphoproteome and the glycoproteome.
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