Agilent, GKE-5002, AdvanceBio Endoglycosidase H

AdvanceBio Endoglycosidase H. Enzyme cleaves within the N,N'-diacetylchitobiose (GlcNAc-GlcNAc) core of high mannose and some hybrid type N-glycans. Includes  5x reaction buffer for GKE-5002 (500 mM Sodium Citrate/Phosphate, pH 5.5, denaturation solution (2% SDS/1M βME, detergent solution (15% detergent solution).

Specifications:
Application: Analysis of biosynthesis of glycoproteins
Concentration: ≥0.15 U/mL
Enzyme Applications: Selective release of high mannose N-linked oligosaccharides from glycopeptides/ glycoproteins. The enzyme has found extensive use in the characterization of glycoproteins, dolichol-linked glycans and the biosynthetic pathway for N-glycosylation.
Enzyme Formulation: 50 mM sodium phosphate pH 7.5, 25 mM EDTA, and 0.05% sodium azide
Enzyme Source: Recombinant gene from Streptomyces plicatus, expressed in E. coli. Enzyme is also known as Endoglycosidase H, mannosyl-glycoprotein endo-beta-N-acetylglucosaminidase.
Enzyme Unit Definition: One unit is defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 60 µmoles of denatured ribonuclease B in 1 hour at 37°C, pH 5.5.
Unit: 30 mU
Volume: 200 µL
pH Optimum: 5.5

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This high-purity endoglycosidase H enzyme is specifically formulated for efficient and selective cleavage of high-mannose and certain hybrid N-linked glycans from glycoproteins. Optimized for reproducibility and robust performance, it supports rapid glycan profiling and structural characterization within biopharmaceutical research and quality control workflows. The enzyme enables precise analysis of glycosylation patterns, aiding in the identification, quantitation, and monitoring of glycan variants throughout candidate development and manufacturing.

Ideal for use in sample preparation protocols prior to liquid chromatography, mass spectrometry, or capillary electrophoresis, this reagent integrates seamlessly with Agilent’s AdvanceBio Glycan Mapping Kits and AdvanceBio LC columns. It facilitates streamlined analysis when paired with Agilent’s 1290 Infinity LC, 6545XT AdvanceBio LC/Q-TOF, and related analytical systems. With its high activity and low background, the product supports rapid deglycosylation even at low enzyme-to-substrate ratios, thereby preserving sample integrity for sensitive downstream analyses.

Comprehensive application guidance ensures reliable integration into established analytical pipelines for monoclonal antibody characterization, biosimilar development, cell line screening, and quality assessment of therapeutic proteins. Compatibility with a broad range of buffer conditions and glycoprotein substrates enhances workflow flexibility in diverse laboratory environments.