Endo F2, Endoglycosidase F2, endo-beta-N-acetylglucosaminidase F2
Source recombinant Elizabethkingia miricola (was Chryseobacterium meningosepticum) in E. Coli
Cleaves all asparagine-linked biantennary oligosaccharides and high mannose (though at a 40X reduced rate) N-glycans from peptides and proteins
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
Specific Activity >20 U/mg
Activity 5 U/ml
Molecular weight 32,000 daltons
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.
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).
Storage Store enzyme at 4˚C.