Reaction Conditions for Endo D (P0742)
ProtocolOptimal incubation times and enzyme concentrations must be determined empirically for a particular substrate. Typical reaction conditions are as follows:
- Combine 10–20 μg of glycoprotein, 1 μl of 10X DTT and H2O (if necessary) to make a 10 μl total reaction volume.
- Denature the glycoprotein by heating the reaction at 55°C for 10 minutes.
- Make a total reaction volume of 20 μl by adding 2 μl 10X GlycoBuffer 2, H2O and 1–5 μl Endo D .
- Incubate the reaction at 37°C for 1 hour.