Preparation of minihepcidins for injection
MW of PR73 = 1,783 Da (but peptide is assumed to be 80% of powder weight) To make enough for twenty 100 nmol doses:- Take 4.4 mg PR73 powder + 700 μl 80% ethanol (4.4 mg instead of 3.5 mg because the peptide is 80% of powder weight)
- Mix solution and inject into SL-220* vial. Heat at 37C until dissolved (a few seconds). Remove cap, transfer 350 μl to each culture tube, and dry in Speedvac until gel is formed
- Add 1 ml injectable quality water per tube for 1 nmol/μl concentration, vortex until gel is dissolved and store at 4C. Inject 100 μl per mouse IP.
Solvent control should be prepared using 700 μl 80% ethanol instead of peptide solution
*Note: Purebright SL-220 is a phospholipid-PEG solubilizer made by NOF Corporation (Japan)
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