I am trying to encapsulate the dye in liposomes and measure pH changes. I have seen a paper on Photochemistry and Photobiology, 2001, 74 (1), 8-13, where they mention that SNARF-1 seems to interact strongly with the membrane. I have tried similar experiments trying to encapsulate the dye in liposomes and change the pH of the bulk solution but it does not look good. They have used DOPC and egg-PC, I have also tried soybean lipids, but the response is quite similar. Has anyone come across that problem or a working protocol for SNARF-1/5F encapsulation in liposomes.
Yes, we used SNARF-1 and also found it tends to stick more to lipid vesicles. When we used protein reconstitution protocols that employ biobeads, we also find that the SNARF-1 sticks to the biobeads, reducing the encapsulation efficiency. It seems SNARF-1 has some hydrophobic character.
Yes, another (annoyed) SNARF-1 user here. There seems to be a tropism towards lipid (or lipophilic) structures. I used SNARF-1 to assess endosomal escape of nanoparticles and liposomes, but I never got anything out of it. I now moved to dual labeled dextran (Rhodamine+Fluorescein). Fluorescein fluorescent is quenched at acidic pH, while Rhodamine fluorescent stays (more or less) constant. Have nice results with this approach.
Many thanks for your input and suggestions. Really appreciate it.
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