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Endoglycosidase F2 [For Glycan Release] (CAT#: CB-P253-K) Datasheet

Product Type
Kit
Description
Selective release of biantennary and high mannose (40-fold reduction) glycans from peptides and proteins.
Endo F2 cleaves N-linked (asparagine-linked) biantennary oligosaccharides from glycoproteins. It can also cut high mannose glycans, but the cutting rate is reduced by 40 times. 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 removes oligosaccharides.
Endoglycosidase F2 is less sensitive to protein conformation than PNGase F, so it is more suitable for natural protein deglycosylation. However, for best results, denaturation of glycoproteins is recommended.
Features
Source: Recombinant Elizabethkingia miricola in E. Coli
EC: 3.2.1.96
Alternative Names: Endo F2, Endoglycosidase F2, endo-β-N-acetylglucosaminidase F2
Specificity:Cleaves all asparagine-linked biantennary oligosaccharides and high mannose (though at a 40X reduced rate) N-glycans from peptides and proteins
Specific Activity:>20 U/mg
Activity:>5 U/mL
Molecular weight: 32,000 daltons
Purity: Endoglycosidase F2 is tested for contaminating protease as follows; 10 µg of denatured BSA is incubated for 24 hours at 37°C with 2 µL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.
Size
0.3 U / 60 µL
Sufficient for up to 60 reactions.
Sensitivity
Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of denatured porcine fibrinogen in 1 minute at 37°C, pH 5.5. Cleavage is monitored by SDS-PAGE (cleaved finbrinogen migrates faster).
Sample
Glycoproteins and glycopeptides containing N-linked glycans
Components
60 µL aliquot of enzyme (0.3 U) in 10 mM sodium acetate 25mM NaCl, pH 4.5
5x Reaction Buffer - 250 mM sodium acetate, pH 4.5
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 4.5
3. Add 2.0 µL of Endo F2. 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

All products and services are for Research Use Only. Do Not use in humans.

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