In the experimental design, the Endo H (MBP Tag) were added to the 20 ul reaction system to investigate the effect of enzymatic digestion on substrate RNase B.
M marker
Lan1 Rnase B 10μg
Lan2 Rnase B 10μg +1U Endo H (MBP Tag)
Endo H (MBP Tag)
Endo H (MBP Tag)
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$96 USD
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Product Details
Product Details
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. |
Background
Endo 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.
Components
Storage 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)
Protocol
1. 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.
Guidelines
Please avoid repeated freeze-thaw cycles.
Unit Definition
One 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.
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Bioactivity
SDS-PAGE
2 μg (R: reducing condition, N: non-reducing condition).
SEC-HPLC
