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