Product Specification| Species | Streptomyces picatus | | Synonyms | Endo-beta-N-acetylglucosaminidase H、Endoglycosidase H、Endo H | | Amino Acid Sequence | / | | Expression System | E.coli | | Molecular Weight | 72kD (Reducing) | | Purity | ＞95% by SDS-PAGE and HPLC | | Conjugation | Unconjugated | | Physical Appearance | Liquid | | Storage Buffer | 20 mM Tris-HCl、50 mM NaCl、5 mM EDTA（pH 7.5 @ 25°C） | | Reconstitution | / | | Stability & Storage | Store at -25 ~ -15℃ for 2 years | | Reference | [1] Wang F , Wang X , Yu X ,et al. High-Level Expression of Endo-β-N-Acetylglucosaminidase H from Streptomyces plicatus in Pichia pastoris and Its Application for the Deglycosylation of Glycoproteins[J].Plos One, 2015, 10.
[2] Maley F, Trimble R B, Tarentino A L,et al. Characterization of glycoproteins and their associated oligosaccharides through the use of endoglycosidases.[J]. Analytical Biochemistry,1989,180(2):195-204. |
BackgroundEndo H is a recombinant glycosidase cloned from Streptomyces plicatus and overexpressed in E.coli. It cleaves the chitobiose core of high-mannose oligosaccharides and a limited number of hybrid oligosaccharides from asparagine-linked glycoproteins, but not complex, oligosaccharides from glycoproteins. ComponentsStorage Solution: 500U/μL EndoH (MBP Tag)、20 mM Tris-HCl、50 mM NaCl、5 mM EDTA (pH 7.5@ 25°C)
10*Denaturing Buffer: 5% SDS、400 mM DTT
10*Reaction Buffer: 500 mM sodium acetate(pH 6 @ 25°C) Protocol1. Combine 1-20 μg of glycoprotein, 1 μl of 10*Denaturing Buffer and H20 (if necessary) to make a 10 μl total reaction volume.
2. Denature glycoprotein by heating raection at 100°C for 10 minutes.
3. Make a total reaction volume of 20 μl by adding 2 μl of 10*Reaction Buffer, H20 and 1-5 μl Endo H (MBP Tag).
4. Incubate reaction at 37°C for 1 hour. GuidelinesPlease avoid repeated freeze-thaw cycles. Unit DefinitionOne unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 µg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10 µl. bio-equip.cn AntBio is a biotechnology group company dedicated to serving life sciences, aiming to help scientists accelerate research and improve work efficiency. AntBio provides comprehensive and high-quality reagent tools for basic research, drug development, and diagnosis, including research grade antibodies, proteins, biochemical reagents, and assay kits. These research tools are widely used in different segments of life science research. The group company currently consists of three brands, Absin, Starter-Bio and UA-Bio.
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