Native and Sequential N-linked Glycan Strategies

For some glycoproteins, no cleavage by PNGase F occurs unless the protein is denatured. For others, some or all of the oligosaccharides can be removed from the native protein after extensive incubation (three days or longer) with PNGase F. PNGase F will remain active under reaction conditions for at least three days, making it suitable for extended incubations of native glycoproteins. Some particular residues,due to their location in the native protein structure, are resistant to PNGase F cleavage and cannot be removed unless the protein isdenatured.

Endoglycosidases F1, F2, and F3 are less sensitive to protein conformation than PNGase F and are more suitable for deglycosylation of native proteins. Endoglycosidase F1 (Endo F1) cleaves asparaginelinked or free high mannose (oligomannose) and hybrid structures, while Endoglycosidase F2 (Endo F2) and Endoglycosidase F3 (Endo F3) have the ability to cleave complex structures. The linkage specificities of Endoglycosidases F1, F2, and F3 suggest a general strategy for deglycosylation of proteins that may remove all classes of N-linked oligosaccharides without denaturing the protein. Asdiscussed previously,complex oligosaccharides can be reduced to the trimannosyldiacetylchitobiose (Man3GlcNAc2) core using neuraminidase, β-galactosidase, N-acetylglucosaminidase, and fucosidases as required. The remaining trimannosyldiacetylchitobiose core structures can be removed with Endoglycosidase F3.
For more information on Glycobiology please browse our online Glycobiology Analysis Manual.

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E9762 Endoglycosidase F1 from Elizabethkingia miricola recombinant, expressed in E. coli, ≥16 U/mg, buffered aqueous solution
E0639 Endoglycosidase F2 from Elizabethkingia miricola recombinant, expressed in E. coli, 20 U/mg
E2264 Endoglycosidase F3 from Elizabethkingia miricola recombinant, expressed in E. coli, 30 U/mg
E7642 Endoglycosidase H from Streptomyces plicatus recombinant, expressed in E. coli, buffered aqueous solution
A0810 Endoglycosidase H from Streptomyces plicatus recombinant, expressed in E. coli, buffered aqueous solution