Suggested In-Solution FASP Digestion Solutions

⦁ AmBic 100 mM: Dissolve 1.40 g in 150 mL ultrapure H₂O. Fill to 200 mL with H₂O in volumetric flask.

⦁ AmBic 50 mM: Prepare from 100 mM AmBic solution with ultrapure H₂O

⦁ Reduction solution: 10 mM DTT in 100 mM AmBic. Make shortly before use.

⦁ Alkylation solution: 15 mM IAA in 100 mM AmBic. Make shortly before use. IAA is light sensitive! Keep it in the dark.

⦁ Stock Trypsin solution 1 µg/µL: Add 100 µL of 50 mM Acetic acid into a 100 µg enzyme vial. Mix by pipetting up and down. Do not vortex! Store 25 µL aliquots at -80°C.

⦁ Trypsin working solution 25 ng/µL: Thaw one tube (enough to digest 8 samples) of stock solution on ice and add 971 µL of 50 mM AmBic.

⦁ Urea 8 M: Dissolve 12.01 g in 50 mM AmBic. Start with a small volume of AmBic since urea increases solution volume. Fill up to 200 mL in volumetric flask.