FAQ: Are Protease Inhibitors acceptable for use with the Protein Deglycosylation Mix reaction?
When a protein is 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 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.