Magnetite MCL Preparation
- Bath Sonicator
- Lipid Mix 
- N-(a-trimethylammonioacetyl)-didodecyl-D-glutamate chloride (TMAG)
- dilauroylphosphatidyl-choline (DLPC)
- dioleoylphosphatidylethanolamine-N-[3-(2-pyridyldithio)-propionate] (PDP-DOPE)
- Lipid marker (?)
- 10 nm magnetite nano-particles
- Specific RDG peptides
- Round bottom flask
- Dissolve the three lipids (in a 1:2:2 molar ratio) into chloroform in the round bottom flask. Do this in a 10-20 mg of lipid/ml organic solvent ratio.
- Remove the organic solvent by either evaporating with dry nitrogen (only for volume of organic solvent less than 1ml) or through rotary evaporation.
- To make sure that all chloroform has been removed from the lipid layer, place the flask on a vacuum pump overnight. Lipid film should coat the bottom of the flask.
- Suspend the magnetite nano-particles in water.
- Hydrate dried lipid layer on the bottom of the flask with nano-particle suspended water.
- Vortex the lipid/water/nano-particle solution for a while.
- Sonicate solution for 30 mins at 28 W in a bath sonicator to reduce to single laminar liposomes.
- Add 2ml of MCLs to the RGD peptides in a .55 peptide to 1 PDP-DOPE molar ratio.
- Gently agitate this solution for 3.5 hours in a solution of 2-morpholinoethanesulfonic acid (MES) buffer (pH 5.0).
- Purify the MCLs.