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Enzymatic Deglycosylation Kit, Premixed cocktail [For Glycan Release] (CAT#: CB-P260-K) Datasheet

Product Type
Kit
Description
Premix the enzyme in a 20 or 100 microliter vial. This allows researchers to quickly remove most glycans (except O-mucin) from glycoproteins under denaturing and natural conditions. By combining these enzymes into an easy-to-use pre-mixed solution, researchers can save time and money for protein deglycosylation. These enzymes can also be provided separately in 20 microliter vials of each enzyme (catalog number CB-P259-K), which gives researchers the flexibility to more fully characterize the glycans linked to their glycoproteins than their enzyme mixtures.
Features
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.
Applications
This kit developed for protein deglycosylation, 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.
Size
20 µL each enzymeEach enzyme; Sufficient for up to 20 reactions.
Sample
Glycoproteins and glycopeptides containing N-linked glycans
Components
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
Handling Advice
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.
Storage
Store enzyme at 4°C.
Shelf Life
Stability: Stable at least 12 months when stored properly. Several days exposure to ambient tempertures will not reduce activity.
Research Use
For research use only. Not for human or drug use

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All products and services are for Research Use Only. Do Not use in humans.

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