How much Endo S should I use to deglycosylate a glycoprotein under native conditions?

When working with native IgG, we recommend using 1 µL of Endo S per 100µg of native IgG. When working with other various glycoproteins (such as Fetuin) we recommend using a modified denaturing protocol, detailed below.

Combine 100 µg of glycoprotein, 1 µL of 400mM DTT and H₂O (if necessary to make a 10 µL total reaction volume.
Denature glycoprotein by heating reaction at 55°C for 10 minutes.
Make a total reaction volume of 20 µL by adding 2 µL 10X GlycoBuffer 1, H₂O and 1-2 µL Endo S.
Incubate reaction at 37°C for 1 hour.

Note: To deglycosylate various glycoproteins, longer incubation times as well as more enzyme may be required. Reactions may be scaled-up linearly to accommodate larger reaction volumes.

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