5.4.1.2. Reduce, alkylate and digest proteins to peptides

  1. For each sample, add 1 µg dithiothreitol (from a stock solution of 1 µg/µl) to 50 µg total protein.
  2. Incubate the solution for 30 min at 37˚C.
  3. Add 5 µg iodoaccetamide (from a stock solution of 5 µg/µl).
  4. Incubate for 20 min at 37˚C.
  5. Add 1 µg trypsin (mass spectrometry-grade, Promega).
  6. Incubate overnight at 37˚C.