FAQ: Are Protease Inhibitors acceptable for use with the Protein Deglycosylation Mix II reaction?

When a protein is reduced or denatured it is more susceptible to cleavage by proteases. For this reason a protease cocktail containing the following can be used with the Protein Deglycosylation Mix II protocol:

  • Aprotinin (10 μg/ml final concentration)
  • Benzamidine (1mM final concentration)
  • Pepstatin (10 μg/ml final concentration)
  • Leupeptin (1μM final concentration)
  • EGTA (1 mM final concentration)
  • EDTA (1 mM final concentration)
  • PMSF (1 mM final concentration)
Make a 1000X concentrated stock of each inhibitor in water; except Pepstatin which should be dissolved in methanol.

Note: PMSF has the ability to modify basic residues on glycoprotein substrates.