Endo F1

远藤 F1

Endo F1 | Endoglycosidase F1 | endo-beta-N-acetylglucosaminidase F1

E489799-20μl

20μl

Specific Activity ＞16 U/mg;Activity ＞17 U/ml

Product Description Endo F1 cleaves Asparagine-linked high mannose and some hybrid oligosaccharides. Core fucosylation reduces the activity by 50 fold. Endoglycosidase F1 will hydrolyze sulfate containing high-mannose chains. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Molecular weight 32,000 daltons Contents 60 µl aliquot of enzyme (1 U) in 20 mM Tris-HCl, pH 7.5 Included with 20 µL and 60 µL pack sizes: 5x Reaction Buffer – 250 mM sodium phosphate, pH 5.5 Specific Activity Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of denatured Ribonuclease B (RNase B) in 1 minute at 37°C, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved RNase B migrates faster). Formulation The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl, pH 7.5 Stability Several days exposure to ambient temperatures will not reduce activity. Stable at least 12 months when stored properly. Specificity Endo F1 cleaves Asparagine-linked high mannose or hybrid oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Quality & Purity Endo F1 is tested for contaminating protease as follows: 10 µg of denatured BSA is incubated at 37°C for 24 hours with 2 µl of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation. The production host strain has been extensively tested and does not produce any detectable glycosidases. Directions for use 1. Add up to 200 µg of glycoprotein to an Eppendorf tube. Adjust to 38 µl final volume with de-ionized water. 2. Add 10 µl 5x Reaction Buffer 5.5 3. Add 2.0 µl of Endo F1 to the reaction. Incubate 1 hour or more at 37°C. Monitor cleavage by SDS-PAGE.

>17 U/ml