Endoglycosidase H (Endo H)
Endoglycosidase H (Endo H)
Cat. No.: SB-G3469-25000U
Size: 25000U
Description
Endoglycosidase H, referred to as Endo H, is derived from the endoglycosidase H of Streptomyces plicatus, which is recombinantly expressed in Escherichia coli, and can specifically cut high mannose structures linked by β-1,4-glycosidic bonds and some hybrid glycoproteins , forming a glycoprotein structure with one N-acetylglucosamine. The cleaved protein still has biological activity and does not affect its function and other follow-up studies. Mainly used to remove N-glycosylation of proteins.
Source: derived from Streptomyces plicatus, recombinantly expressed in Escherichia coli;
Definition of enzyme activity: at 37°C, the amount of enzyme required to remove more than 95% of carbohydrates in 10 μg of denatured RNase B for 1 hour is defined as an enzyme activity unit;
Purity and concentration: SDS-PAGE detection purity > 95%; endogenous nucleic acid residue < 1 pg/μL (qPCR detection); 500 U/μL;
Inactivation or inhibition: Heat at 75°C for 10 minutes to inactivate;
Enzyme storage buffer: 20 mM Tris-HCl, 50 mM NaCl, 5 mM EDTA, 50% Glycerol, pH 7.5;
10х Denaturing Buffer: 5% SDS, 400 mM DTT;
10х Native Buffer: 400 mM sodium acetate, pH 6.0;
Figure 1. The deglycosylation effect of the substrate RNase B treated with Endo H. After the substrate RNase B was denatured, 10 μg was added to different enzyme amounts of Endo H (0, 10, 1, 0.5, 0.1 U), incubated at 37°C for 1 h, and detected by PAGE electrophoresis.
Storage and transportation
Transport in wet ice packs; store at -20°C, valid for 12 months.
Product Contents
| Component Number | Component | G3469 |
| G3469-1 | Endoglycosidase H (Endo H) | 50 μL |
| G3469-2 | 10х Denaturing Buffer | 1mL |
| G3469-3 | 10х Native Buffer | 1mL |
| Product Manual | 1 copy | |
Steps
1. Protein deglycosylation under denaturing conditions:
Add 1 μL 10х Denaturing Buffer and dH2O to the 1-20 μg protein sample to the 10 μL reaction system, and incubate the system at 100°C for 10 min. Then add 2 μL 10х Native Buffer, 1-5 μL Endo H and appropriate amount of dH2O to the 20 μL reaction system, and incubate at 37°C for 1 h.
2. Protein deglycosylation under non-denaturing conditions:
Add 2 μL 10х Native Buffer, 2-5 μL Endo H and appropriate amount of dH2O to 1-20 μg protein sample to 20 μL reaction system, and incubate at 37°C for 4-24 h. (Protein deglycosylation under non-denaturing conditions requires longer reaction times or more enzymes)
Precautions
1. SDS will not affect the activity of the enzyme.
2. The easiest way to assess the degree of protein deglycosylation is by SDS-PAGE gel electrophoresis.
3. All enzymes should be stored in an ice box and stored at -20°C immediately after use.
4. For your safety and health, please wear a lab coat and disposable gloves for operation.