2D Analysis of Phosphoproteins & Phosphopeptides

Pro-Q® Diamond phosphoprotein stains are ideal for analysis of phosphorylation of a single protein or an entire proteome. Pro-Q® Diamond phosphoprotein gel stain can be used with both 1-D and 2-D gels and is fully compatible with mass spectrometry. In addition to the Pro-Q® Diamond phosphoprotein gel stain, some of our kits include SYPRO® Ruby total-protein stain and PeppermintStick™ phosphoprotein molecular weight standards to serve as extra controls in your experiments. We also offer the Pro-Q® Diamond Phosphoprotein Blot Stain Kit, suitable for detecting phosphoproteins on PVDF or nitrocellulose membranes.

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A 2-D gel was stained first with Pro-Q® Diamond phosphoprotein gel stain and subsequently with SYPRO® Ruby protein gel stain. In this section of the gel, the computer-generated overlay of the two staining patterns reveals the phosphoproteins (magenta), and nonphosphorylated proteins (green). Where the signals overlap, the spots appear gray.

The Pro-Q® Diamond phosphoprotein/phosphopeptide microarray stain directly detects phosphorylated proteins or peptides on microarrays — no antibodies or radioisotopes are required. You can use the stain to identify kinase targets on protein arrays, or make your own kinase identification array with peptide substrates. This product works optimally on microarrays with polyacrylamide gel surfaces.

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Casein kinase assay on a kinase target peptide microarray using the Pro-Q® Diamond Phosphopeptide/Phosphoprotein Microarray Stain Kit.

With the EZQ™ Phosphoprotein and Phosphopeptide Quantitation Kits, you get easy and fast quantitation of up to 96 samples at a time. The innovative format of this assay assures accurate quantitation from just 1 mL of sample in the presence of detergents, reducing agents and urea buffers containing up to 1% of carrier ampholytes. These kits are ideal for measuring phosphatase and kinase activity and for monitoring relative phosphoprotein or phosphopeptide concentrations during chromatography or after IEF fractionation of protein samples.

• EZQ™ Phosphoprotein and Phosphopeptide Quantitation Kit
• EZQ™ Phosphoprotein and Phosphopeptide Quantitation Kit

EZQ™ Phosphoprotein Quantitation Kit.

The Captivate™ Microscale Phosphopeptide Isolation Kit provides a highly selective method for isolating phosphopeptides from complex solutions without using antibodies. The technology is ideal for the isolation of phosphorylated peptides prior to liquid chromatography and/or mass spectrometry separations. This streamlined method is suitable for single samples or high-throughput environments.

Selective quantitative isolation of phosphopeptides using the Captivate™ Microscale Phosphopeptide Isolation Kit. A standard mixture of nine peptides containing approximately 550 pmol of total phosphopeptide was incubated with 400 mg of Captivate ferrofluid phosphopeptide-binding reagent. The starting mixture, supernatant, and eluate were then analyzed by HPLC.

Our Pro-Q® Diamond LC Phosphopeptide Detection Kit selectively tags phosphopeptides with a fluorescent marker, providing a highly sensitive method for selective detection of phosphopeptides over non-phosphopeptides. At the same time, the phosphopeptide retention time is altered, allowing for easy identification and purification. The kit is ideal for the development of front-end HPLC phosphopeptide separations prior to mass spectrometry.

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The Pro-Q® Diamond LC phosphopeptide detection reagent binds to the phosphopeptides in a tryptic digest of -casein, which are revealed using excitation at 550 nm during HPLC fractionation.