Thermo Scientific™ Pierce™ Phosphoprotein Staining Kit
Detect phosphoserine and phosphothreonine residues in phosphorylated proteins with this in-gel, colorimetric phosphoprotein staining kit.
Manufacturer: Thermo Scientific™ PI24552
Thermo Scientific™ Pierce Phosphoprotein Staining Kit is for specific, colorimetric, in-gel detection of phosphoserine and phosphothreonine residues in phosphorylated proteins.The Pierce Phosphoprotein Staining Kit is a colorimetric stain to specifically detect proteins separated by polyacrylamide gel electrophoresis (SDS-PAGE) that are phosphorylated at serine and/or threonine residues. Staining is achieved by first hydrolyzing the phosphoprotein-phosphosphoester linkage using 0.5N NaOH in the presence of calcium ions. The gel containing the newly formed insoluble calcium phosphate is then treated with ammonium molybdate in dilute nitric acid. The resulting insoluble nitrophospho-molybdate complex is finally stained with a solution of the basic dye, methyl green. The reagents in this kit hydrolyze the phosphoester linkage of phosphoserine and phosphothreonine. Phosphotyrosine is not hydrolyzed and cannot be detected with this kit. Although the staining method is not sufficiently sensitive to detect subtle changes in phosphorylation of low-abundance proteins, the kit provides a convenient system for assessing purification and phosphorylation status of phosphoproteins during preparative workflows.
Specific for pSer and pThr phosphoproteins – for use with SDS-PAGE to detect abundant serine- and threonine-phosphorylated proteins in the sample
Easy-to-follow protocol – produces results in approx. three hours, yielding phosphoproteins that are identified as green to green-blue bands
Qualitative staining – detection limit depends on the extent of phosphorylation and protein size and abundance; detects 40 to 80ng phosvitin and 80 to 160ng beta-casein per band
Controls included – complete kit includes one positive control protein (phosvitin) and one negative control protein (soybean trypsin inhibitor) for method validation
Compatible with double-staining – phosphoprotein-stained gels can be secondarily stained with Thermo Scientific GelCode Blue Stain Reagent (Part No. 24590) for total protein profiling
Seven solutions (3L total), plus two vials of lyophilized protein (1mg each)
Dry incubator heated to 65°C.
Evaluate the progress of a phosphoprotein purification process; identify approximate molecular weights of most abundant phosphoproteins in a sample
|Pierce Phosphoprotein Control Set|
|STI negative control protein, 1mg, Phosvitin positive control protein, 1mg|
|Phosphoprotein control lanes in at least 10 gels|
|Vials of lyophilized phosvitin (40kDa phosphoprotein) and soybean trypsin inhibitor (STI, 20kDa), 1mg each|
|2 Protein Set|