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Endoglycosidase F1 [For Glycan Release] (CAT#: CB-P252-K) Datasheet

Product Type
Kit
Description
Endo F1 cleaves high mannose and some hybrid N-glycans from peptides and proteins.
Endo F1 cleaves asparagine-linked high mannose and some hybrid oligosaccharides. Core fucosylation reduces the activity by 50 times. Endoglycosidase F1 will hydrolyze sulfates containing high mannose chains. It cleaves between the two N-acetylglucosamine residues in the diacetyl chitobiose core of the oligosaccharide to produce a truncated sugar molecule, one of which remains in the asparagus Amide on. In contrast, PNGase F completely eliminated oligosaccharides.
Features
Source: Recombinant Elizabethkingia miricola in E. Coli
EC: 3.2.1.96
Alternative Names: Endo F1, Endoglycosidase F1, endo-β-N-acetylglucosaminidase F
Endo F1 Specificity: Cleaves all asparagine-linked high mannose and some hybrid oligosaccharides
Specific Activity:>16 U/mg
Activity:>17 U/mL
Molecular weight: 32,000 daltons
Size
1 U / 60 µL
Sensitivity
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).
Sample
Glycoproteins and glycopeptides containing N-linked glycans
Components
60 µL aliquot of enzyme (1 U) in 20 mM Tris-HCl, pH 7.5
5x Reaction Buffer - 250 mM sodium phosphate, pH 5.5
Handling Advice
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. Incubate 1 hour at 37°C.
Storage
Store enzyme at 4°C.
Shelf Life
Stability: Stable at least 12 months when stored properly. Several days exposure to ambient tempertures will not reduce activity.
Research Use
For research use only. Not for human or drug use

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