The QBT™ Fatty Acid Uptake Assay is a homogeneous assay amenable to high-throughput screening. The kits deliver pre-optimized, fluorescence-based formulations to expedite assay development and screening of fatty acid transporters.
Conventional protocols for monitoring fatty acid uptake using radioactivity often require cell lysis and processing at very low temperature, making them expensive, slow and unsuitable for high throughput screening. Other fluorescence-based protocols generally require the use of low throughput fluorescence activated cell sorter (FACS) instrumentation, or require cell washing, which can severely compromise the integrity of the fragile adipocytes used in these assays.
The QBT Fatty Acid Uptake Assay uses a BODIPY®-dodecanoic acid fluorescent fatty acid analog coupled with Molecular Devices proprietary quench technology. The BODIPY label provides an ideal long chain fatty acid analog that behaves much like natural fatty acids and is a known substrate for fatty acid transporters.
- 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
- Replaces cumbersome and time-consuming fluorescent and radioactive assay protocols
- The elimination of radioactive compounds reduces disposal costs and eliminates safety risks associated with radioactive assays
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 the 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.
- 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
For more details, please refer to the QBT Fatty Acid Uptake Assay Kit product insert, which may be accessed through the Molecular Devices Knowledge Base.
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.
BODIPY is a registered trademark of Life Technologies Corporation.
Fibroblasts vs. adipocytes
Fibroblasts vs. adipocytes; insulin dose-response
Leptin inhibitory effect
QBT Fatty Acid Uptake Assay Kit is compatible with these systems:
- FLIPR Tetra High Throughput Cellular Screening System
- FlexStation 3 Benchtop Multi-Mode Microplate Reader
- SpectraMax i3 Multi-Mode Detection Platform
- SpectraMax M5e/M5/M4/M3/M2e Microplate Readers
- SpectraMax Paradigm Multi-Mode Microplate Reader
- Gemini EM Microplate Reader
For more information, view the assay kits & instrument compatibility table.
|Product Name||Description||Part Number|
QBT Fatty Acid Uptake Assay Kit
1 vial = 1 plate x 2 vials each
1 vial = 1 plate x 10 vials
1 vial = 5 plates x 10 vials
Compatible with 96- and 384-well plate formats.