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O-Glycosidase [For Glycan Release] (CAT#: CB-P256-K) Datasheet

Product Type
Kit
Features
Specificity:
O-glycosidase leaves only unsubstituted Gal-β(1-3)GalNAc-α disaccharides attached to the serine or threonine residues of glycoproteins or glycopeptides.

Substitutions such as sialic acid, galactose, fucose or N-acetylglucosamine must first be removed with the appropriate exoglycosidase prior to treatment with Endo-O-Glycosidase. At minimum, a sialadase such as Sialadase Au α(2-3,6,8,9), cat. number CB-P257-K, is almost always required to remove sialic acids.

Source: Recombinant Streptococcus pneumoniae in E.Coli

EC: 3.2.1.97

Specific Activity:>12 U/mg

Activity:>1.25 U/mL

Molecular weight: 180,000 daltons

pH range: 5-7, optimum 5.0

Purity: O-Glycosidase is tested for contaminating protease as follows; 10 µg of denatured BSA is incubated for 24 hours at 37°C with 2 µL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.
Size
75 mU / 60 µL; Sufficient for up to 60 reactions.
Sensitivity
Specific Activity: Defined as the amount of enzyme required to produce 1 µmole of p-nitrophenol (pNP) in 1 minute at 37°C, pH 5.0 from p-nitrophenyl-2-acetamido-2-deoxy-3-O-(β-D-galactopyranosyl)-α-D-galactopyranoside.
Sample
Glycoproteins and glycopeptides containing N-linked glycans
Components
60 µL aliquot of enzyme (75 mU) in 50 mM sodium phosphate, pH 7.5.
5x Reaction Buffer 5.0 (250 mM sodium phosphate, pH 5.0)
Handling Advice
1. Add up to 100 µg of glycoprotein to a tube.
2. Add de-ionized water to a total of 13 µL.
3. Add 4 µL of 5x Reaction Buffer 5.0.
4. Add 1 µL of Sialadase Au α(2-3,6,8,9), cat. number CB-P257-K.
5. Add 2 µL of O-Glycosidase.
6. Incubate for 1 hour at 37°C.
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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