Endo F1,UA BIOSCIENCE_specification/price/image

Product Detail
Company Profile

Product Specification

Species	Elizabethkingia meningoseptica
Synonyms	Endo-beta-N-acetylglucosaminidase F1; EndoF1; Di-N-acetylchitobi-osyl beta-N-acetylglucosaminidase F1; Endoglycosidase F1; Mannosyl-glycoprotein endo-beta-N-acetyl-glucosaminidase F1
Expression System	E.coli
Molecular Weight	33kDa (Reducing)
Purity	＞95% by SDS-PAGE&HPLC
Tag	His Tag
Physical Appearance	Liquid
Storage Buffer	20mM Tris, pH7.5 (@ 25°C)
Stability & Storage	Store at -25 ~ -15℃ for 2 years
Reference	1. Van Roey P, Rao V, Plummer T H, et al. Crystal structure of endo-beta-N-acetylglucosaminidase F1, an alpha/beta-barrel enzyme adapted for a complex substrate. [J].Biochemistry, 1994, 33(47):13989-96. 2. Elder J H, Alexander S. endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins[J]. Proc Natl Acad Sci U S A, 1982, 79(15):4540-4544.

Background

Endo-beta-N-acetylglucosaminidaseF1(EndoF1) is an endoglyco-sidasee, secreted by Elizabethkingia meningoseptica, that cleaves aspa-raginelinked oligosaccharides after the first N-acetylglucosamine residue. The enzyme is selective for high-mannose oligosaccharide chains. Glycosylation of the core fucoidan of the heterodimeric structure reduces the cleavage rate of EndoF1 by more than EndoF1 hydrolyzes sulfate containing high mannose chains. EndoF1 can be used under natural or non-denatured deglycosylation conditions. The enzyme is suitable for deglycosylation of glycoproteins under natural conditions and is suitable for a variety of glycomics applications, proteomics and mass spectrometry applications.

Components

UA070121: 10U/µL EndoF1 in 20mM Tris, pH7.5 (@ 25°C)
10*Reaction buffer:500 mM Sodium phosphate, pH 5.5 (@ 25°C)

Protocol

1.Take up to 200µg of glycoprotein or glycopeptide sample, adjust to 43µl with purified water;
2.Add 5µl of 10× reaction buffer;
3.Add 2µl EndoF1 to a total volume of 50µl, mix gently;
4. React for 1 hour at 37°C.

Guidelines

1. For different glycoprotein samples, the optimal enzyme concentration and reaction time need to be mapped experimentally;

2. Higher enzyme concentrations increase the digestion efficiency of individual glycoproteins and need to be optimized accordingly;

3. SDS and DTT will limit enzyme activity;

4. The final concentration of the enzyme after dissolution will be stored at -20℃ for storage after dispensing as needed.

Unit Definition

One unit is defined as the amount of enzyme that more than 95% of carbohydrates were removed from denatured 5 μg RNase B at 37°C, reacting 1 hour, in a 10 μL reaction system.

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.