Molecular Devices introduces FA uptake assay
assay kit that addresses the need for a single step, homogenous in
vitro assay for fatty acid uptake for use in the identification of
potential new drugs via high-throughput screening.
The QBT fatty acid uptake assay kit aims to provide researchers with a greater insight into the defects that occur during fatty acid metabolism. This defect has been linked with several pathological states including insulin desensitisation, Type 2 diabetes, obesity and cardiovascular disease.
The kit is a significant advancement over conventional assays, which use radioisotopes. Conventional protocols utilizing radioactivity often require cell lysis and processing at very low temperature, making them expensive, slow and not suitable for high throughput screening. Fluorescence-based protocols generally require the use of low throughput Fluorescence Activated Cell Sorter (FACS) instrumentation, or require cell washing, which can compromise fragile adipocyte cells.
The homogeneous format makes the kit less labour intensive to run. With the single step mix-and-read procedure, the cells can be incubated with the reagents and are stable for up to several hours. Rapid analysis of the cells can be followed with the detection on any bottom-read fluorescence plate reader. The assay is suited for high throughput screening applications in both 96-well and 384-well formats.
Uniquely, the kit employs a BODIPY -dodecanoic acid fluorescent fatty acid analog. The BODIPY label provides a long chain fatty acid analog that behaves much like natural fatty acids. It becomes activated by acyl-CoA attachment, which is incorporated into di-and triglycerides. These accumulate in intracellular lipid droplets. In addition, the BODIPY analog is a known substrate for fatty acid transporters since its uptake by adipocytes can be competed by non-labelled fatty acids.
Fatty acids are utilized for diverse cellular processes including mitochondria oxidation, membrane synthesis and energy storage. Pathologically increased intracellular fatty acid concentrations can cause cellular apoptosis and have been linked to insulin desensitization, Type 2 diabetes, obesity and cardiovascular disease.
All of these intracellular processes are dependent upon fatty acids traversing the plasma membrane to get into the cell. Therefore, understanding the Fatty Acid Transport Proteins (FATP) that regulate this process is of great importance in biomedical research and drug discovery.
A spokeswoman for Molecular Devices said: "Since this assay uses a fluorescence-based readout rather than a radioactivity-based one, it eliminates the safety risks associated with conventional radiolabel assays, in addition to being faster and easier to perform."
