FAQ: What is a typical reaction protocol for Endo D?

We recommend using 1 µL of Endo D per 20µg of paucimannose N-linked glycans.
  1. Combine 10-20 µg of glycoprotein, 1 µL of 10X DTT and H2O (if necessary to make a 10 µL total reaction volume.
  2. Denature glycoprotein by heating reaction at 95°C for 3 minutes.
  3. Make a total reaction volume of 20 µL by adding 2 µL 10X GlycoBuffer 2, H2O and 1 µL Endo D.
  4. Incubate reaction at 37°C for 1 hour.
Note: To deglycosylate various glycoproteins under native conditions, 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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Endo D