Enzymatic DeGlycoMx Kit
This kit developed for protein deglycosylation includes our DeGlycoMx, a premixed cocktail of the enzymes required to remove all N-linked oligosaccharides and most O-linked sugars from 0.5 mg of glycoprotein, via 10 reactions of up to 50 micrograms of protein per reaction.
The kit is easy-to-use and effective: add 2 µL of the DeGlycoMx enzyme to your denatured sample and the included buffer and incubate for 3 hours at 37°C.
View product documentation: Specsheet / CofA / MSDS
- Amount of Enzyme
- 20 µL each enzyme
Kit includes enzyme plus reaction buffers.
Sufficient for up to 20 reactions.
The enzymes are premixed in either one 20 or 100 microliter vial. This allows researchers to quickly remove most all glycans (with the exception of O-Linked mucins) from their glycoprotein in both denatured and native conditions. By combining the enzymes into one easy-to-use premixed solution, researchers save time and money in the pursuit of protein deglycosylation. The enzymes can also be supplied individually in 20 microliter vials of each enzyme (KE-DG01) allowing the researcher to the flexibility to characterize the glycans attached to their glycoprotein more fully than with our mixture of enzymes.
The enzymes are provided as one 20 µL premixed cocktail including: PNGase F (Elizabethkingia meningosepticum), O-Glycosidase (Streptococcus pneumoniae), Sialidase (Arthrobacter ureafaciens), β-Galactosidase (Streptococcus pneumoniae), Glucosaminidase (Streptococcus pneumoniae).
5x Reaction buffer - 200 µL
Denaturation Solution - 100 µL
Triton X - 100 µL
The Enzymatic DeGlycoMx Kit will remove all N-linked oligosaccharides and many O-linked oligosaccharides from glycoproteins. Protein deglycosylation for N-linked glycans (Asparganine-linked) is performed using the enzyme PNGase F. In addition, all Serine or Threonine linked (O-linked) Gal-(β1-3)-GalNAc-(α1) and all sialic acid substituted Gal-(β1-3)-GalNAc-(α1) will be removed using the combination of Sialidase and O-Glycosidase. The addition of β-Galactosidase and Glucosaminidase will assist in the deglycosylation of larger O-link structures.
1. Mix 10 µL of reaction buffer with up to 50 µg of glycoprotein in 33 µL distilled water in a 1.5 µL tube.
2. Add 2.5 µL denaturation solution. Mix gently and place in boiling water bath for 5 minutes. Chill on ice.
3. Add 2.5 µL of Triton-X.
4. Add 2 µLs of the DeGlycoMx enzyme cocktail. Incubate for 3 hours at 37°C.
Note: Denaturation increases the rate of enzyme digestion up to 10 fold. If denaturation is not desired omit step 2-3, add with 5 µL of distilled water and increase incubation time up to 24 hours.
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