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PNGase F catalyzes de-N-glycosylation in a domestic microwave.

Analytical biochemistry (2012-04-21)
Hui Zhou, Andrew C Briscoe, John W Froehlich, Richard S Lee
ABSTRACT

Common de-N-glycosylation protocols usually require a lengthy incubation time. Although pressure cycling technology or scientific microwave reactors can accelerate this enzyme reaction, they may not be easily accessible. In this brief report, we employed an alternative strategy using a standard domestic microwave oven to perform the de-N-glycosylation. Model glycoproteins (bovine RNase B, bovine fetuin, and human IgG) and a complex mixture from human plasma were fully deglycosylated in 20 min, without any apparent adverse affects on the glycans or protein backbones. This new method provides a simple and inexpensive solution to achieve rapid de-N-glycosylation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
PNGase F from Elizabethkingia meningoseptica, BioReagent, ≥95% (SDS-PAGE), for proteomics
Sigma-Aldrich
PNGase F from Elizabethkingia miricola, buffered aqueous solution
Sigma-Aldrich
Glycopeptidase A from almonds, buffered aqueous glycerol solution, ≥0.05 unit/mL
Sigma-Aldrich
PNGase F from Elizabethkingia meningoseptica, ready-to-use solution, recombinant, expressed in E. coli
Sigma-Aldrich
PNGase F from Elizabethkingia meningoseptica, recombinant, expressed in E. coli, set of 100 units nanomolar unit
Sigma-Aldrich
PNGase F from Elizabethkingia meningoseptica, lyophilized powder, recombinant, expressed in E. coli