Fatty acids are essential dietary nutrients consisting of a carboxylic acid moiety linked to a long aliphatic hydrocarbon chain. They are the fundamental building blocks of more complex lipids such as those that form cellular membranes. In the blood, long-chain fatty acids travel bound to serum albumin and are transported into cells by members of the SLC27 family of transmembrane transporter proteins. Uptake of long-chain free fatty acids into adipocytes is essential for regulating circulating lipid levels and cellular lipid metabolism. Abnormalities in adipocyte fatty acid uptake and lipid metabolism are major contributing factors to obesity, diabetes/metabolic syndrome, cardiovascular disorders and nonalcoholic fatty liver disease (NAFLD). As diet and exercise consistently fail to achieve any lasting results in combating obesity, pharmacological targeting of fatty acid uptake is being investigated as a potential treatment strategy. The Fatty Acid Uptake Assay Kit enables rapid determination of real-time fatty acid uptake kinetics in live cells expressing fatty acid transporter proteins. The assay uses a highly fluorescent long-chain fatty acid analogue (Ex/Em = 488/523 nm) that acts as a substrate for transmembrane fatty acid transporters (FATPs) and accumulates in intracellular lipid droplets. A proprietary non-toxic membrane-impermeable quenching agent eliminates any fluorescence arising from the extracellular space, ensuring specific measurement of intracellular fatty acid accumulation without requiring any tedious wash steps. The assay is highly sensitive, has a simple mix-and-read protocol and is high-throughput adaptable for screening of fatty acid uptake-modulating test compounds or treatment conditions. The kit contains a complete set of reagents sufficient for performing 100 reactions in a 96-well plate format. Sample types: adherent cells expressing transmembrane fatty acid transporters, differentiated 3t3-l1 adipocytes, differentiated 3t3-l1 heterologous cells
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Characterization of fatty acid uptake kinetics in live cells.. Screening and characterization of drugs and test compounds for modulation of fatty acid uptake.