Direct approach for estimation of protein carbohydrate content.

The Thermo Scientific Pierce Glycoprotein Carbohydrate Estimation Kit enables the amount of protein glycosylation to be measured as the percent of total purified protein mass.

The Glycoprotein Carbohydrate Estimation Kit includes six purified glycoprotein standards and the required assay reagents to effectively estimate the amount of oxidizable glycosylation (percent carbohydrate by weight) of purified protein samples. Sugar groups in the glycoprotein sample are first oxidized with sodium meta-periodate to produce detectable aldehyde groups. Then the sample is reacted with the colorimetric Glycoprotein Detection Reagent. Absorbance at 550nm of the resulting purple reaction product is measured with a spectrophotometer or plate reader. Finally, the carbohydrate content in the glycoprotein is calculated by comparison to results from the five glycoprotein standards that are included in the kit.

Highlights:

• Qualitative – easily identifies purified proteins as glycoproteins or samples as contaminated with sugars
• Semi-quantitative – estimates the percent carbohydrate content (w/w) of purified glycoprotein by comparison to the included set of glycoprotein standards
• Simple procedure – completed in less than 75 minutes; instructions include microplate and test tube protocols
• Adaptable – standard curve format allows for the design of alternative tests for aldehydes and carbohydrate components

Applications:

• Determine if a purified polyclonal antibody or other protein is glycosylated before attempting to perform carbohydrate-based conjugation or immobilization chemistries with hydrazide reagents
• Characterize and classify purified protein fractions from an affinity procedure
• Assess sugar and carbohydrate contamination in non-glycosylated protein samples

Related Products and Resources:

Glycoprotein Carbohydrate Estimation Kit FAQ
Protein Assay Selection Guide
Pierce Glycoprotein Staining Kit
Krypton Fluorescent Glycoprotein Staining Kit
Glycoprotein Isolation Kits