Hi Gustavo,

1) You are right, not too many options are available with 12:0 chain length, though you might try with head-group labelled 14:0, such as 14:0 NBD PE from Avanti (or any similar).

2) Dil might work for you, though the 18:0 chains look a bit long. But you can try first without the protein to see if the incorporation reaches the level needed for quantification.

3) There is a shorter Dil, e.g. D-383 or DiIC12(3)

4) You might try to quatify the lost utilizing the protein's fluorescence, e.g. lookinf for the tryptophan signal.

Best,

Karoly

You don't necessarily have to include a fluorescent lipid in the liposomes to measure the amount of lipid recovered. You could do an assay for lipid. Since you used PC, you could do an assay for choline. A kit for this is available from Wako. Another approach is to assay the amount of lipid using an extrinsic lipid-sensitive fluorescent dye and standards with known amounts of the same lipid.

Adam is right, you can try the choline assay, or run a simple TLC plate with appropriate standards and spray the plate with primulin.

Best,

Karoly

Dear Karoly and Adam,

Thanks for your valuable suggestions!

From all of other possibilities that don't include fluorescent lipid and DiI, which one do you think has the best balance between price and easiness, considering that I may have to buy more reagents than I actually need? I ask because I already have 16:0 Liss Rhod PE and DiO available, so if I'm going to change the strategy, it has to make up.

Best regards,

Gustavo