FAQ: Can glycosidases be used on whole cells?

NEB glycosidases can be used in conditions that are compatible with whole or live cells. In order to keep cells viable, non-denaturing conditions are typically recommended for use with whole or live cells. Serum, and other common culture media components, do not seem to have a deleterious effect on NEB glycosidases. The pH of the reaction should be kept between pH 6.0 – 8.0 (notice that some enzymes have optimal pH below this range). It is recommended to include a protease inhibitor cocktail to prevent protein degradation. Any cocktail may be used, however we recommend avoiding one which includes PMSF, as PMSF can modify basic residues.

Although glycosidases are generally active under these conditions, their expected action might not be complete. As a substrate becomes more complex (from protein to whole cells in culture media) accessibility is impaired, and conditions might not be optimal for maximal activity. One will have to determine empirically the amount of glycosidase needed to remove the desired carbohydrate. As a recommendation, use more enzyme and longer incubation times.