Enzymatic DeGlycoMx Kit

Kit includes the QA-Bio DeGlycoMx™ enzyme cocktail and the reagents required to remove all N-linked oligosaccharides and many O-linked sugars from 1 mg of glycoprotein in one 3 hour incubation. Enough reagents are included for ten deglycosylation reactions. The enzymes are premixed in one 20 microlitre vial. This kit 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 deglycosylation..
Includes one 20 microlitre vial including the following enzymes in solution: PNGase F (Chryseobacterium meningosepticum) O-Glycosidase (Streptococcus pneumoniae) Sialidase (Arthrobacter ureafaciens) ß-Galactosidase (Streptococcus pneumoniae) Glucosaminidase (Streptococcus pneumonia)	Other Supplied Reagents: Reaction buffer Denaturation Solution Triton X
Specificity The Enzymatic DeGlycoMx™ Kit will remove all N-linked oligosaccharides and many O-linked oligosaccharides from glycoproteins. N-links (Asparganine-linked) are removed using the enzyme PNGase F. In addition, all Serine or Threonine linked (O-linked) Gal-(b1-3)-GalNAc-(a1) and all sialic acid substituted Gal-(b1-3)-GalNAc-(a1) will be removed using the combination of Sialidase and O-Glycosidase. The addition of ß-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.