Carboxy-SNARF 1 (5/6-mixture)

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Carboxy-SNARF 1 (5/6-mixture) for fluorescent staining of liposomes^[1]
1. Preparation of working solution
Dissolve Carboxy-SNARF 1 (5/6-mixture) in HEPES/50 mM KCl buffer (pH7.5) to prepare a 0.1 mM Carboxy-SNARF 1 (5/6-mixture) working solution.
2. Specific staining steps
1) Weigh 10 mg of phospholipid, which can be dioleoyl L-phosphatidylcholine (DOPC) or egg yolk lecithin (EYL).
2) Dissolve the phospholipid in 0.5 mL chloroform and place it in an Eppendorf tube.
3) Dry the solution under a stream of nitrogen to form a lipid film on the inner wall of the tube.
4) Place the tube in a vacuum for 10 min.
5) Add 0.5 mL of 0.1 mM Carboxy-SNARF 1 (5/6-mixture) working solution to the vial containing the lipid film.
6) Mix the solution thoroughly using a vortex shaker to form multilamellar vesicles.
7) Freeze the liposome suspension in liquid nitrogen.
8) After thawing, prepare large unilamellar vesicles (LUVs) with a diameter of 400 nm by extrusion using a LipoFast extrusion device.
9) Remove the dye outside the liposomes by gel filtration. Centrifuge at 600 g for 5 min in pH 7.5 buffer using a 5 cm Sephadex G25 column.
10) The stained LUVs are ready for fluorescence measurement.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.