33kDa (Reducing)
20mM Tris, pH7.5 (@ 25°C)
Store at -25 ~ -15℃ for 2 years
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.
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.
UA070121: 10U/µL EndoF1 in 20mM Tris, pH7.5 (@ 25°C)
10*Reaction buffer:500 mM Sodium phosphate, pH 5.5 (@ 25°C)
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.
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.
Different amounts of EndoF1 were utilized to degrade denatured RNaseB under the reaction conditions of 37°C, 1h.
M: marker
Lane1: EndoF1
Lane2: Denatured RNaseB
Lane3: EndoF1/ denatured RNase B=
1/200
Lane4: EndoF1/ denatured RNase B=
1/400(1U)
Lane5: EndoF1/ denatured RNase B=
1/800
Lane6: EndoF1/ denatured RNase B=
1/1600
2μg (R: reducing condition, N: non-reducing condition).
SEC≥95%
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