Endo F1 (Endoglycosidase F1) is a recombinant glycosidase expressed and purified from E.coli which carries the Chryseobacterium meningosepticum gene.Endo F1, like Endo H, cleaves between the N-acetylglucosamine residues of the chitobiose core of N-linked glycans, leaving one N-acetyl glucosamine residue attached to the asparagine.Endo F1 has a molecular weight of ~33.6 kDa.The workable pH range is between 5.0 to 6.0, with the optimal pH at 5.5.
Substrate Specificity- High mannose N-linked oligosaccharides- Hybrid N-linked oligosaccharides Product Component- Endo F1 : 100 ㎕ (500 unit/㎕)- 10x Reaction Buffer : 500 mM sodium citrate, pH 5.5. Unit Definition- One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 µg of denatured RNase B for 1 hr at 37°C in a total reaction volume of 10 ㎕. Specific Activity- 500 unit/㎕, 2,000 unit/㎍ Storage Buffer- 20 mM Tris-HCl, pH 7.5, 50 mM NaCl, 5 mM EDTA, 50% glycerol. Endo F1 Reaction Condition- 10 µg of RNase B are denatured with 0.5% SDS, 1% β-mercaptoethanol at 100°C for 10 min. After the addition of Reaction Buffer to final 1x, Endo F1 are added and the reaction mix is incubated for 1 hr at 37°C. Separation of reaction products is analyzed by SDS-PAGE (cleaved RNase B migrates faster). The activity of Endo F1 is not affected by addition of 0.5% SDS. For a native glycoprotein, 5-fold more enzyme and longer incubation time will be required. Storage Temperature- Store at -20℃. QC Tests
- Activity, SDS-PAGE purity, performance tests .