Endoglycosidase F3 [For Glycan Release] (CAT#: CB-P254-K) Datasheet

Product Type
Kit
Description
Selectively release triantennarry and fucosylated biantennary N-glycans N-glycans from peptides and proteins.
Endo F3 cleaves free or asparagine-linked tritactose or α(1-6) fucosylated bicontact oligosaccharides, as well as the core structure of tricarbamylchitobiose. Non-fucosylated biantennary glycans will also be cleaved, but the decomposition 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 in which one N-acetylglucosamine residue remains in the asparagine on. In contrast, PNGase F completely removes oligosaccharides. Alpha (1-3) fucosylation will inhibit enzyme activity. It has no activity on oligomannose and hybrid molecules.
Features
Source: Recombinant Elizabethkingia miricola in E. Coli
EC: 3.2.1.96
Alternative Names: Endo F3, Endoglycosidase F3, endo-β-N-acetylglucosaminidase F
Specific Activity:
Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of porcine fibrinogen in 1 minute at 37°C, pH 4.5. Cleavage is monitored by SDS-PAGE (cleaved fibrinogen migrates faster).
Purity: Endoglycosidase F3 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.
Specificity:Cleaves all asparagine-linked triantennary or α(1-6) fucosylated biantennary oligosaccharides, as well as triamannosyl chitobiose core structures. α(1-3) fucosylation will inhibit enzymatic activity. Nonfucosylated biantennary glycans will also be cleaved, but at a 40x reduced rate. (Note: The recombinant version is not glycosylated, which may result in properties differing from the native protein.)
Specific Activity:>25 U/mg
Activity:>5 U/mL
Molecular weight: 30,000 daltons
Size
0.33 U / 60 µL
Sensitivity
Defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 micromole of porcine fibrinogen in 1 minute at 37°C, pH 4.5. Cleavage is monitored by SDS-PAGE (cleaved fibrinogen migrates faster).
Sample
Glycoproteins and glycopeptides containing N-linked glycans
Components
60 µL aliquot of enzyme (0.33 U) in 20 mM Tris-HCl, pH 7.5
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
4. Add 2.0 µL of Endo F3. 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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