16 May 2023 1 4K Report

Hi,

I'm trying to learn how to do liposome uptake assays but i can't get understand why some papers do "counter-flow" liposome uptake assays while most of the papers don't.

By liposome uptake assay, i'm referring to experiment in which membrane proteins are reconstituted into liposomes, and the uptake of substrates by the proteoliposomes are measured.

In 'normal' liposome assay, you just incubate the proteoliposome in an external buffer that has your radioactive substrate.

In the 'counter-flow' liposome assays, people pre-load proteoliposome with unlabeled substrate and incubate the proteoliposome in external buffer with radioactive substrate at a much lower concentration compared to the concentration of unlabeled substrate inside the liposome.

What's the advantage of doing the counter flow assay over the default one?

I would really appreciate your help.

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