Jhdavis::PeptideCleavage

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  1. Transfer from block to glass scintillation vials
    1. Take block out of peptide sysnthesis hoot (hard to lift and easy to break)
    2. In hood, remove cam from well
    3. Using p1000 with cut tim, use 1 mL EtOH to transfer resin to glass scint. vial.
    4. Pipet off extra ethanol -> EtOH waste
    5. Let dry o/n.
  1. Cleavage
    1. There is a choice of scavengers (EDT seems good for everything) - see choice list in the hood.
    2. For peptides containing met, 94% TFA, 1 % TIS - thioisopropylene, 2.5% water, 2.5% EDT) - need 5 mL per peptide
    3. ~2 hrs. with shaking (put ether on ice during this time)
    4. Filter off resin - using glass wool in the pipette (pre-chill pipette/peptide)
  1. Wash
    1. Wash 4x in total with ether
    2. Spin 3000k for 10 min (swinging bucket with top sealed)
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