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Endoglycosidase H [For Glycan Release] (CAT#: CB-P255-K) Datasheet

Product Type
Kit
Description
Endo H cleaves asparagine-linked hybrid or high mannose oligosaccharides, but does not cleave complex oligosaccharides.
Endo H cleaves asparagine-linked hybrid or high mannose oligosaccharides, but does not cleave complex oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetyl chitobiose core of the oligosaccharide to produce a truncated sugar molecule in which one N-acetylglucosamine residue remains in the asparagine on. In contrast, PNGase F completely removes oligosaccharides. Detergents and heat denaturation can increase the cleavage rate of certain glycoproteins.
Features
Source: recombinant from Streptomyces plicatus in E.Coli
EC: 3.2.1.96
Alternative Names: Endo H, endo-β-N-acetylglucosaminidase H, Endoglycosidase H
Specific Activity: One unit of Endo H activity is defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 µmole of denatured Ribonuclease B. Cleavage is monitored by SDS-PAGE (cleaved Ribonuclease B migrates faster).
Specific Activity:>40 U/mg
Activity:>5 U/mL

Molecular weight: 29,000 daltons
pH range: 5-6, optimum 5.5
Purity: Endoglycosidase H 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
0.3 U / 60 µL
Sufficient for up to 60 reactions.
Sample
Glycoproteins and glycopeptides containing N-linked glycans
Components
60 µL aliquot of enzyme (300 mU) in 20 mM Tris-HCl, 25 mM NaCl, 1 mM EDTA (pH 7.5).
200 µL 5x Reaction Buffer 5.5 (250 mM sodium phosphate, pH 5.5)
Denaturation Solution - 2% SDS, 1 M Beta-mercaptoethanol
Handling Advice
1. Add up to 200 µg of glycoprotein to Eppendorf tube. Adjust to 37.5 µL final volume with deionized water.
2. Add 10 µL 5X Endo H Buffer and 2.5 µL of Denaturation Solution (SDS/-ME). Heat at 100°C for 5 minutes.
3. Cool, and then add 2.0 µL of Endo H to the reaction. Incubate 3 hours 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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