The Thermo Scientific Pierce Glycoprotein Staining Kit is a convenient, fast and sensitive kit for staining glycosylated proteins in polyacrylamide gels using the periodic acid-Schiff (PAS) method.
The kit provides three essential reagents and a complete protocol to specifically stain glycosylated proteins (glycoproteins) that have been separated by polyacrylamide gel electrophoresis (PAGE). A gel or membrane containing separated proteins is treated with a periodate solution, which oxidizes cis-diol sugar groups in glycoproteins. The resulting aldehyde groups are detected through the formation of Schiff-base bonds with a reagent that produces magenta bands. Traditionally called the periodic acid-Schiff (PAS) reagent method, this staining technique is made easier by the convenient packaging of essential reagents in one kit with easy-to-follow instructions. The kit includes positive and negative control proteins.
- Specific for glycoproteins – periodic acid-Schiff (PAS) method specifically detects glycosylated proteins having sialic acid and other oxidizable carbohydrate groups
- Optimized protocol – the traditional three-reagent method has been streamlined to achieve optimal staining in less than two hours (half the time required for other staining methods)
- Colorimetric – glycoproteins are detected as magenta bands against a faint pink or colorless gel background
- Qualitative staining – detection limit depends on the extent of glycosylation and protein size and abundance; detects 0.625ng avidin and 0.16µg horseradish peroxidase per band
- Controls included – complete kit includes positive and negative control proteins (HRP and STI, respectively).
- Gels or membranes – effective for use with polyacrylamide mini gels or nitrocellulose membranes
- Three reagents in 250mL bottles (1 solution and 2 dry powders to which water must be added), plus two vials of lyophilized proteins (1mg each)
|Protocol summary for the Thermo Scientific Pierce Glycoprotein Staining Kit.
To use the Pierce Glycoprotein Staining Kit, simply treat the gel or membrane containing separated proteins treated with the provided periodate-based Oxidation Reagent. This treatment oxidizes cis-diol sugar groups in glycoproteins. The resulting aldehyde groups are reacted with the Pierce Glycoprotein Stain Reagent to form Schiff-base bonds that result in the production of magenta bands of the stain. Traditionally called the periodate-acid-Schiff (PAS) reagent method, this staining technique is made easier by the convenient packaging of essential reagents in one kit with easy-to-follow instructions.
|Sensitive, specific staining of glycosylated proteins with the Thermo Scientific Pierce Glycoprotein Staining Kit. Identical protein samples were electrophoresed in identical polyacrylamide gels and then stained with the Pierce Glycoprotein Staining Kit (left) or the Thermo Scientific GelCode Blue Stain Reagent for total protein detection (right). Only the 5 or 6 proteins containing different amounts of glycosylation are detected with the Glycoprotein Stain. The left lane in each panel is prestained MW marker.
- Misenheimer, T.M., (2001). J. Biol. Chem. 276, 45882-45887.
- Pio, R., et al. (2001). J. Biol. Chem. 276, 12292-12300.
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