Preparation of bOG-DOPG mixed micelles

 Preparation of bOG-DOPG mixed micellesContributor:Date: November 22, 1993 Materials: All glassware must be acid washed, rinsed thoroughly with water then rinsed with acetone and dried. Redistill acetone.Recrystallizing BOG:1) Add 5 grams of Octyl-b-D-glucopyranoside (bOG = Calbiochem #494459) to 20 ml acetone.2) Heat mixture at 40°C to completely dissolve the bOG.3) Filter solution through a Pyrex® scintered glass funnel.4) Slowly add 100 ml diethyl ether to the filtered solution and re-heat if necessary (if precipitation appears in the solution).5) Place in -20°C freezer overnight (or longer) to allow crystals to form.6) Pour solution containing recrystallized bOG through a scintered glass funnel.7) Wash crystals with 500 ml ice cold diethyl ether.8) Completely dry crystals and scrape out of funnel.9) Weigh crystals and store as powder in the -20 °C freezer.Preparing mixed micelles:10) Aliquot 0.97 ml of 20 mg/ml L-a-dioleoyl-phosphatidylglycerol (DOPG sodium salt = Avanti #840475) into Pyrex® screw-cap tubes.--> NOTE: DOPG stock solution should be 20 mg/ml or 27 mM in chloroform.11) Dry the DOPG as a thin film around the bottom of the glass tube.--> NOTE: If the DOPG is dried as a pellet at the bottom of the tube it is more difficult (takes more time) to dissolve in aqueous solution.12) Make up a 7.5% solution of bOG in water and add 1 ml to each tube containing dried DOPG.13) Sonicate mixture until the DOPG is completely dissolved.--> Do 4 minute bursts with 5 minute rests in between. Depending on how the DOPG was dried in the tube, dissolving the DOPG completely will take 2 to 4 hours.