Benefits of the QBT™ Fatty Acid Uptake Assay Kit

·               Replaces cumbersome and time-consuming radioactive assays, reducing disposal costs and eliminating safety risks associated with radioactive assays.

·               One-step, homogeneous assay format enables high-throughput screening.

·               Fluorescent assay with proprietary quench technology allows users to monitor real-time uptake kinetics in live cells.

Assay Principle

QBT Fatty Acid Uptake Assay Kit includes a loading buffer that contains fluorescent fatty acid analog, along with a quenching dye to greatly reduce fluorescence in the extracellular space.  When loading buffer is added to cells, transport of fluorescent fatty analog into the cells results in an increase in fluorescence signal that is monitored in real time using a bottom-reading fluorescence microplate reader.  Inhibition of fatty acid transport by test compounds results in reduced fluorescent signal relative to controls.

Workflow

·               Prepare differentiated adipocytes from 3T3-L1 fibroblasts, or other FATP-expressing cell type as desired (for details on cell preparation, please refer to the QBT Fatty Acid Uptake Assay Kit product insert).

·               Plate cells so that a confluent monolayer forms.

·               Serum deprive the cells if necessary.

·               Treat cells with test compounds.

·               Add the recommended volume of loading buffer to wells.

·               Transfer the plates immediately to a fluorescence microplate reader and set the microplate reader to read wells every 20 seconds for 30-60 minutes.

Patented technology

QBT Assay Kits from Molecular Devices employ a quenching dye to reduce background fluorescence and improve the signal-to-noise ratio. The patented quench technology is offered to drug discovery and life science researchers exclusively by Molecular Devices, through the purchase of QBT Assay Kits.

Quenching technology is covered under U.S. Patent 6,420,183 and EPO Patent 0 906 572.

To place an order, call your local Molecular Devices office or Molecular Devices distributor.