I do patch clamp (and Ca2+ imaging) and usually apply DAGs-DMSO in external buffer through the perfusion.
Now I want to see if my ion channels will respond to cholesterol.
I prepared Cholesterol-MbCD solution according to the protocol. I incubated and perfused cells with Cholesterol-MbCD but cells did not respond to the treatment.
(Could be I made a mistake when was preparing Chol-MbCD or did not store it properly)
The problem is that the preparation of Cholesterol-MbCD is very long. SO I would like to know if I can simplify it.
I could not find any paper where cholesterol dissolved in DMSO and then applied in patch camp experiments (or Ca2+ imaging).
Does anyone have an experience with cholesterol application in patch clamp (or Ca2+ imaging)?
And what is the best way to prepare cholesterol for the experiments?
Is it possible to apply Cholesterol-DMSO to the cells?
Thank you so much for your help!
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