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.

Contents:
Ludger Enzymatic DeGlycoMx - Kit contents
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).
Also includes:
5x Reaction buffer - 200 µL
Denaturation Solution - 100 µL
Triton X - 100 µL

Specificity:
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.

Directions:
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.