| Species | Streptomyces picatus |
| Expression System | E.coli |
| Molecular Weight | 30kD (Reducing) |
| Conjugation | Unconjugated |
| Tag | His Tag |
| Physical Appearance | Liquid |
| Storage Buffer | 20 mM Tris-HCl、50 mM NaCl、5 mM EDTA（pH 7.5 @ 25°C) |
| Stability & Storage | Store at -25 ~ -15℃for 2 years |
| Reference |
[1] Wang F , Wang X , Yu X ,et al. High-LevelExpression of Endo-β-N-AcetylglucosaminidaseH from Streptomyces plicatus in Pichia pastoris and Its Application for theDeglycosylation of Glycoproteins[J].Plos One, 2015, 10. [2] Maley F, Trimble R B,Tarentino A L,et al. Characterization of glycoproteins and their associatedoligosaccharides through the use of endoglycosidases.[J]. Analytical Biochemistry,1989,180(2):195-204. |
Endo H is a recombinant glycosidase cloned fromStreptomyces plicatus and overexpressed in E.coli. It cleaves thechitobiose core of high-mannose oligosaccharides and a limited number of hybridoligosaccharides from asparagine-linked glycoproteins, but not complex,oligosaccharides from glycoproteins.
StorageSolution: 500U/μL EndoH、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 sodiumacetate(pH 6 @ 25°C)
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°Cfor 10 minutes.
3.Make a total reaction volume of 20 μlby adding 2 μl of 10*ReactionBuffer, H20and 1-5 μl Endo H.
4. Incubate reaction at 37°C for 1 hour.
Please avoid repeated freeze-thaw cycles.
In the experimental design, 125 U, 25 U, 5 U, 1 U and 0.2 U enzymes were added to the 20 ul reaction system to investigate the effect of Endo H enzyme on the enzymatic digestion of substrate RNase B.
M marker
Lan1 blank
Lan2 Rnase B 10μg
Lan3 Rnase B 10μg +125U Endo H
Lan4 Rnase B 10μg+ 25U Endo H
Lan5 Rnase B 10μg+ 5U Endo H
Lan6 Rnase B 10μg+ 1U Endo H
Lan7 Rnase B 10μg+ 0.2U Endo H
2μg (R: reducing
condition, N: non-reducing condition).
