Abstract
It has been shown that the Dole procedure extracts as much free fatty acids from rat liver microsomes as does homogenization in chloroform-methanol, 2:1. If microsomal phospholipids in the heptane phase from the Dole procedure are removed with silicic acid, microsomes give a background in the 63Ni assay for free fatty acids equivalent to about 10 nmol of fatty acid/mg of microsomal protein. Only about 2% of this background is due to endogenous free fatty acids. Thus, use of the Dole extraction procedure, “clean-up” of the hexane phase with silicic acid, and the 63Ni assay allow determination of the hydrolysis of 5% or less of microsomal phospholipids using only small amounts of material (0.5–1.0 mg of microsomal protein). This procedure was found to be useful in following the breakdown of microsomal phospholipids by phospholipase A 2.
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