I have read so many papers on vitD3 acting as differentiation agent. I tried this and found that I got differentiation only once when I freshly dissolved calcitriol in ethanol and thereafter I never observed even after increasing concentration. It's seems to be highly air sensitive. Do I need to make aliquots in anaerobic glove box? Or could you suggest me the right way to use it?
When I used to work with calcitriol, everything was always done in low lighting. Calcitriol is very light-sensitive. We also prepared working stocks in EtOH and foil-wrapped many small aliquots. I don't think we used an aliquot more than once or twice because of its instability. When cultures were treated, we always used low lighting (i.e., no light from windows or direct ceiling light). Also make sure there is serum in the medium (or BSA for serum-free medium) to assure the calcitriol will stay in solution and not end up sticking to the sides of your medium container. Hope this helps.
@terence Mcguire:- I did same. I made small aliquots in very low light condition and used EtOH only as solvent. Calcitriol worked only once when i freshly dissolved, subsequently used aliquots did not work, seems its highly air sensitive.
I cannot say whether it is air-sensitive or not. We did not seem to have that problem (at least after being dissolved in EtOH). You might want to check if your cells have down-regulated the vitamin D receptor since your first experiment.
Terence McGuire - Hi Terence. Sorry to stretch back to an old thread; do you recall what you used to store your working stock aliquots in? And do you recall using low-retention tips? Both to combat the 'stickiness' of VitD metabolites. Thanks
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