FFAR4 (GPR120) Reporter Assay

FFAR4/GPR120是一种G蛋白偶联受体，可作用于长链脂肪酸，如DHA和EPAω-3脂肪酸。它参与调节脂肪细胞的葡萄糖摄取，并介导巨噬细胞的抗炎活性。Cayman的FFAR4(GPR120)报告基因检测试剂盒由96孔板组成，该板涂有转染复合物，其中包含FFAR4短同工型(FFAR4S)的DNA构建体、将Gαq活化信号引导至Gαs通路的工程G蛋白和cAMP反应元件调节的SEAP报告基因(FFAR4反向转染条板)。在转染复合物上生长的细胞将在细胞表面表达FFAR4S，并在细胞内表达重组G蛋白。激动剂与FFAR4S的结合会启动信号转导级联，导致SEAP表达，并分泌到细胞培养基中。刺激后16-24小时收集培养基的等分试样，并在添加试剂盒中提供的基于发光的碱性磷酸酶底物后测量SEAP活性。该试剂盒易于使用，可轻松应用于高通量筛选调节FFAR4S激活的治疗化合物。试剂盒中包含一种已知的FFAR4激动剂GSK137647A1，可用作阳性对照。该试剂盒提供足够的试剂，可使用三个随附的白色测定板在三个时间点测量SEAP活性。FFAR4/GPR120 is a G protein-coupled receptor for long chain fatty acids such as DHA and EPA ω-3 fatty acids. It is involved in the regulation of glucose uptake in adipocytes and mediates anti-inflammatory activity in macrophages. Cayman's FFAR4 (GPR120) Reporter Assay Kit consists of a 96-well plate coated with a transfection complex containing DNA constructs for the short isoform of FFAR4 (FFAR4S), an engineered G protein that directs Gαq activation signals to the Gαs pathway, and a cAMP response element-regulated SEAP reporter (FFAR4 reverse transfection strip plate). Cells grown on the transfection complex will express FFAR4S at the cell surface and the recombinant G protein inside the cell. Binding of agonists to FFAR4S initiates a signal transduction cascade resulting in expression of SEAP which is secreted into the cell culture medium. Aliquots of culture medium are collected 16-24 hours after stimulation and SEAP activity is measured following addition of a luminescence-based alkaline phosphatase substrate provided in the kit. The kit is easy to use and can be readily applied to high-throughput screening for therapeutic compounds regulating activation of FFAR4S. A known FFAR4 agonist, GSK137647A1, is included in the kit for use as a positive control. The kit provides sufficient reagent to measure SEAP activity at three time points using the three included white assay plates.分子式分子量