In Vitro: Protocol 1) Combine 2 μg substrate and 50 mM ammonium bicarbonate pH 8.5 to make a 10 μL total reaction volume. 2) Add α-Lytic Protease in a 1:20 – 1:100 (α-Lytic Protease : substrate) ratio by mass. Note: For example, add 20-100ng α-Lytic Protease to 2 μg substrate protein. 3) Incubate at 37℃ for 1 hour. Note: longer incubations (18 hours) or more enzyme may be required, especially at lower temperatures, for complete digestion.