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Enzymatic DeGlycoMx Kit

This kit 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 as PDF

Part Number: KE-DGMX

$330.00

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)
  • Beta-Galactosidase (Streptococcus pneumoniae)
  • Glucosaminidase (Streptococcus pneumonia)

Other Supplied Reagents:

  • Reaction buffer
  • Denaturation Solution
  • Triton X
Deglycosylation Image

Specificity
The Enzymatic DeGlycoMx䋢 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 Beta-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.

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