脂質代謝
プロメガ脂質代謝アッセイは、グリセロール、トリグリセライド、コレステロールおよびコレステロールエステルを検出および定量するための高感度でシンプルな方法を提供します。これらのアッセイは、生物学的プロセスの中でも、脂肪分解および脂肪生成の定量化に使用することができます。脂質代謝アッセイは、他のプロメガ代謝アッセイでも採用されているコア生物発光技術をベースにしています。
脂質代謝アッセイは、単層または3D構造の培養細胞、組織、細胞培養培地、血漿、血清など、多くの生体サンプルに適応できます。脂質代謝アッセイで使用される細胞タイプにには、単層および3D培養した肝細胞および脂肪細胞も含まれます。このプロトコルでは、面倒な有機抽出ステップが不要なので、サンプル調製が飛躍的に簡素化されます。また、このアッセイは広範な直線性を示すとともに感度も高いためサンプルを希釈する回数を低減し、脂質代謝産物の微小な変化をとらえる定量能力を備えています。
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Introduction to Lipid Metabolism Assays
Options for lipid metabolism assays include bioluminescent plate-reader based assays to detect glycerol, triglyceride, cholesterol and cholesterol ester levels in biological samples. These assays are fast and easy to use, as they require no organic extraction and only two incubation steps (at either 37°C or room temperature), and provide quantitative results with the use of a standard curve. Unlike staining and other methods that require organic extraction, these bioluminescent assays are simple to implement for laboratories that are new to lipid metabolism research.
Lipids perform many roles in an organism, including signaling functions, acting as components of biological membranes and as a form of energy storage. Understanding changes in levels of lipids in biological samples such as cell culture samples, cultured 3D cell structures, tissues, and plasma or serum samples is a use of lipid metabolism assays.
Alteration of lipid metabolism results in a variety of disorders. Steatosis refers to the aberrant accumulation of lipids in tissues. Steatosis and the measurement of abnormal lipid levels in tissues is relevant to metabolic disorder research, including diabetes, obesity and liver disorders. Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), two liver disorders drastically increasing in global prevalence, are key targets of many drug discovery programs. Lipid metabolism assays are useful tools in the study of these disorders.
Lipolysis is the process by which cells hydrolyze one molecule of triglyceride to three molecules of fatty acids and one molecule of glycerol. An increase in lipolysis can be measured as an increase in free glycerol levels in cell culture samples using a glycerol assay.
Lipogenesis is the creation of triglyceride from one molecule of glycerol and three molecules of fatty acid, and it is a way in which cells store energy. An increase in lipogenesis can be measured as the quantification of intracellular triglycerides using a triglyceride assay. Triglyceride assays quantitate the levels of triglyceride by determining the total level of glycerol, the level of free glycerol and the resulting difference is the level of triglyceride.
Cholesterol is an essential component of cellular membranes and is required to produce hormones. However, an increased level of circulating cholesterol is associated with increased risk of cardiovascular disease and atherosclerosis. Cholesterol is transported in plasma in the form of cholesterol ester, in low-density lipoproteins (LDL) or high-density lipoproteins (HDL). One molecule of cholesterol ester is converted to one molecule of cholesterol via the action of the enzyme cholesterol esterase. Cholesterol assays are used to measure total cholesterol (including cholesterol esters) and free cholesterol levels, in a variety of sample types.
