Lipid Extraction questions - Lipid (Sep/05/2005 )
I'm not sure if this is the appropriate place for lipid related questions.
I would like to begin looking at amounts of ceramide in my cells. I have noticed that the protocols state that the extracted lipids should be 'dried under nitrogen'. We do not have a visidry manifold (as recommended by one publication), and as we do not intend to to do a significant amount of work on ceramide it may not be worth investing in one. Is there an alternative? Is it essential to dry under nitrogen? Is this to do with it being solvent evaporation?
Secondly, every protocol has specified the use of glass tubes. Is this due to a problem with lipids binding to plastic? Our scintillation counter manual only recomends the used of plastic tubes, do you think this would be a problem?
I would be very grateful for any help!
If you don't dry under nitrogen, the samples become oxidated.. this messes up results.
Yes, you must use glass, this is an organic extraction.. it will melt plastic tubes.
You can do your scintillation counting in plastic tubes.
u can use butylated hydroxytoulene to prevent lipid oxidation...