Typical Reaction Conditions for Endo F3 Protocol (P0771)
- Combine 20 µg of glycoprotein, 1 µl of 10X GlycoBuffer 4 and H2O (if necessary) to make a 10 µl total reaction volume.
- Add 1 µl Endo F3
- Incubate reaction at 37°C for 1 hour.
The reaction may be scaled-up linearly to accommodate large amounts of glycoprotein and larger reaction volumes.
The optimal pH for the cleavage of fucosylated-biantennary (i.e. porcine fibrinogen) and triantennary glycans (i.e. fetuin) is pH 4.5 (10X GlycoBuffer 4). However, fucosylated-biantennary glycans can also be cleaved at pH 6.0 (10X GlycoBuffer 3).
The enzyme should be used under non-denaturing conditions. Detergents, such as SDS, inhibit Endo F3 activity.