1. Run 1D or 2D protein gel. 3-50 pmole of the protein to be sequenced may be needed. Concentrating the sample before loading if necessary (blowing stream of N₂, Speed Vac, or precipitation).

2. Transfer the proteins to the PVDF membrane using a full immersion tank and 10 mM CAPS(3-cyclohexylamino-1-propanesulfonic acid), 10% methanol, pH 11 as the transfering buffer instead of the normal buffer containing Tris and SDS.

3. Staining the proteins on the PVDF membrane with Ponceau S or Amido Black.

4. Cut the desired band.