Suggested In-Solution 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

⦁ 150 mM IAA (IodoAcetAmide) - Add 27.7 mg of IAA to 1 mL of 50 mM AmBic

⦁ 45 mM DTT (DiThioThreitol) - Add 6.95 mg of DTT to 1 mL of 50 mM AmBic

⦁ 5% Trifluoroacetic acid (TFA) - Add 5 mL to 100 mL nanpure H₂O

⦁ RapiGest SF: Add 500 μL of AmBic to a 1 mg vial of RapiGest

⦁ 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-Lys-C working solution 25 ng/µL: Thaw one tube (enough to digest 24 samples) of stock solution on ice and add 971 µL of 50 mM AmBic.

⦁ Wetting solution I: 100% LC-MS grade acetonitrile (ACN)

⦁ Wetting solution II: 50% LC-MS grade ACN

⦁ Elution solution: 0.1% TFA in 50% ACN

⦁ Equilibration & Washing solutions: 0.5% TFA in 5% ACN

⦁ Sample solution: 2% TFA in 20% ACN