Digestion Recipies

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Suggested In-Gel 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

⦁ Destain solution: Prepare fresh by mixing ACN with 100 mM AmBic (1:1, vol/vol).

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

⦁ Alkylation solution: 55 mM IAA in 100 mM AmBic. Make shortly before use. IAA is light sensitive! Keep it in dark.

⦁ Trypsin stock solution 100 ng/µL: Add 200 µL 50 mM Acetic acid into a 20 µg trypsin vial. Mix by pipetting up and down in tip. Do not vortex! Store 30 µL aliquots at -80°C.

⦁ Trypsin working solution 15 ng/µL: Thaw one tube of stock solution on ice and add 170 µL of 50 mM AmBic.

⦁ Stop solution: 5% (v:v) formic acid in 50% (v:v) ACN.

⦁ Elution solution A: 1% (v:v) formic acid in 60% (v:v) MeOH.

⦁ Elution solution B: 1% (v:v) formic acid in 50% (v:v) ACN.

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

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

⦁ Equilibration & Sample solutions: 0.1% TFA in HPLC water

⦁ Washing solution: 0.1% TFA in 5% LC-MS grade Methanol

⦁ Elution solution: 0.1% TFA in 50% ACN