β(1-2,3,4,6)-N-acetylglucosaminidase [For Glycan Release] (CAT#: CB-P268-K) Datasheet

Product Type
Kit
Description
N-acetylglucosaminidase cleaves all non-reducing terminal β-linked N-acetylglucosamine residues from complex carbohydrates and glycoproteins.

The cleavage rate of the different bonds of GlcNAc on two-antenna, three-antenna, and four-antenna oligosaccharides largely depends on the steric hindrance of adjacent residues. Β(1-2) GlcNAc residues linked to β(1-3)-linked mannose are cleaved at the highest rate, while β(1-2) residues linked to β(1-6)-linked mannose) GlcNAc residues were cleaved at the highest rate in 2000. All three oligosaccharides have the lowest ratios. β(1-6)GlcNAc residues (if present) are removed at the second highest rate, while β(1-4)GlcNAc is removed at the third highest rate. On the three-antenna structure, the residue is removed at the second highest rate. The bisected β(1-4)GlcNAc linked to β-linked mannose severely hinders the cleavage of other GlcNAc residues-cleavage requires high concentrations of enzymes and prolonged incubation time.
Features
Source: Recombinant from Streptococcus pneumoniae in E. Coli.

EC: 3.2.1.52

Alternate Names: β-N-acetylglucosaminidase, N-acetyl-β-d-glycosaminide N-acetylglucosaminohydrolase, glucosaminidase, hexosaminidase

Specific Activity:>80 U/mg
Activity:>50 U/mL

Molecular weight: ~140,000 daltons

pH range: 5-7, optimum 5.0

Specificity: Cleaves all non-reducing terminal β-linked N-acetylglucosamine. Bisecting GlcNAc slows the reaction.

Specific Activity Assay: 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-β-D-N-acetyl-glucosaminide
Applications
N-acetylglucosaminidase cleaves all non-reducing terminal β-linked N-acetylglucosamine residues from complex carbohydrates and glycoproteins.
Size
2.4 U / 60 µL; Sufficient for up to 60 reactions.
Sample
Glycoproteins and glycopeptides containing N-linked glycans
Components
N-acetylglucosaminidase in 20 mM Tris-HCl, 25 mM NaCl, pH 7.5
5x Reaction Buffer 250 mM sodium phosphate, pH 5.0
Handling Advice
1. Add up to 100 µg of glycoprotein or 1 nmole of oligosaccharide to a tube.
2. Adjust to 14 µL final volume with de-ionized water.
3. Add 4 µL 5x reaction buffer (pH 5.0).
4. Add 2 µL of N-acetylglucosaminidase.
5. Incubate 3 hours at 37°C.
Storage
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.

ONLINE INQUIRY

Creative Biolabs has established a team of customer support scientists ready to discuss ADCC/CDC optimization strategies, antibody production, bioinformatics analysis and other molecular biology/biotechnology issues.

  • *
  • *
  • *
USA

UK