Small scale cleave peptide from the resin

  1. Place ~30resin beads in a 1.5mL Eppendorf tube, and add 1 drop of cleavage cocktail K.
  2. Place the tube at room temperature for 1 hour.
  3. Blow away TFA with a glass pipette in the chemistry hood.
  4. Add 200uL diethyl ether and mix.
  5. Desktop centrifuge the peptide at top speed for 1 min.
  6. Dump the supernatant and you should be able to see a small white precipitate at the bottom of the EP tube.
  7. Dry the tube in air for about 1 min and dissolve the peptide in 50uL 50% ACN/water.
  8. Spin again and take 10uL of the supernatant by avoiding the resin beads, and mixture with another 100uL 50% ACN/water.
  9. Inject in ESI to check purity.

Cleavage cocktail K: TFA/thioanisole/water/phenol/EDT (90 : 2.5 : 2.5 : 2.5 : 2.5 v/v)