I am planning experiment in which I would like to use detergent based lysis to prepare cellular lysates for cholesterol quantiation by the cholesterol oxidase method. I am aware that endogenous peroxides etc may interfere with the quantification and will deal with it as suggested here: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2952578/?report=printable#bib9
I have seen that most methods employ an organic solvent method for the isolation of lipids. However, I am curious as to whether initial lysis could be performed using a detergent method with subsequent lipid isolation being performed on a lysate aliquot by the hexane:isopropanol or other method for use in an enzymatic cholesterol quantitation assay.
try using more gentle detergents such as n-butnol. It has been validated in enzymatic quantification of triacylglycerol.
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