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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

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

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