184.108.40.206. Reduce, alkylate and digest proteins to peptides
- For each sample, add 1 µg dithiothreitol (from a stock solution of 1 µg/µl) to 50 µg total protein.
- Incubate the solution for 30 min at 37˚C.
- Add 5 µg iodoaccetamide (from a stock solution of 5 µg/µl).
- Incubate for 20 min at 37˚C.
- Add 1 µg trypsin (mass spectrometry-grade, Promega).
- Incubate overnight at 37˚C.