Preparation of minihepcidins for injection

MW of PR73 = 1,783 Da (but peptide is assumed to be 80% of powder weight) 1 nmole = 1733 ng = 1.7 μg pure peptide or 2.2 μg 80% peptide. We have used doses between 10 and 100 nmoles (100 nmoles is a very high dose that causes profound hypoferremia).

50 nmol PR73 dose per mouse = ~0.11 mg PR73 and 1.5 mg solvent (SL220).

Use 100 μl volume for subcutaneous or IP injection.

To make enough for (for example) twenty 50 nmol doses:

Take 2.2 mg PR73 powder + 350 μl 80% ethanol (2.2 mg instead of 1.73 mg because the peptide is 80% of powder weight)
Take 30 mg SL220 + 350 μl 80% ethanol, warm up to 37 C to dissolve (a few seconds)
Mix PR73 and SL220, and warm up again if it is difficult to dissolve
Split liquid between two tubes, and dry in Speedvac until gel is formed
Add 1 ml injectable quality water per tube for 0.5 nmol/μl concentration, vortex until gel is dissolved and store at 4C. Inject 100 μl per mouse SQ or IP.

Solvent control should be prepared using 350 μl 80% ethanol instead of peptide solution

*Note: Purebright SL-220 is a phospholipid-PEG solubilizer made by NOF Corporation (Japan)), and is now called DSPE-020CN.
USA contact: https://www.nofamerica.com/store
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