Home Protocols Reaction conditions for Simultaneous Digestion of IgG with IdeZ Protease (IgG-specific) and PNGase F (fragmentation and deglycosylation) (P0770)

Reaction conditions for Simultaneous Digestion of IgG with IdeZ Protease (IgG-specific) and PNGase F (fragmentation and deglycosylation) (P0770)

Optimal PNGase F units may vary for a particular antibody. Typical reaction conditions are as follows:
  1. Combine up to 50 μg of human IgG and H2O to a volume of 25 μl.
  2. Add 2.5 μl of 10X GlycoBuffer 2.
  3. Add 1 μl (500 units) of PNGase F Glycerol Free (NEB# P0709) and 1 µl (80 units) of IdeZ Protease (IgG-specific).
  4. Incubate 2 hours at 37°C.