Schmidt John A
Department of Biology, Villanova University, Mendel Hall Rm G63B, 800 Lancaster Avenue, Villanova, PA, 19085, USA.
Methods Mol Biol. 2016;1496:187-95. doi: 10.1007/978-1-4939-6463-5_15.
Determining the abundance of phospholipids and neutral lipids in cellular membranes is paramount to understanding their biological functions. Many lipid-modifying enzymes have yet to be characterized due to limitations in substrate-product measurements and purification of membrane-bound enzymes. The method described here uses radiolabeled phospholipid substrates and cell-purified organelles to quantify phospholipid metabolism using thin-layer chromatography. This assay has the benefits of being specific and adaptable for numerous applications and systems.
确定细胞膜中磷脂和中性脂质的丰度对于理解其生物学功能至关重要。由于底物-产物测量和膜结合酶纯化方面的限制,许多脂质修饰酶尚未得到表征。本文所述方法使用放射性标记的磷脂底物和细胞纯化的细胞器,通过薄层色谱法定量磷脂代谢。该测定法具有特异性强且适用于多种应用和系统的优点。