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

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

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