QBT Fatty Acid Uptake Assay Kit
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 our 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
Latest Resources
Data of QBT Fatty Acid Uptake Assay Kit
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Fibroblast vs. adipocytes
3T3 L1 adipocytes were plated at 50,000 cells/well in 100 µl of DMEM/FBS in a 96-well plate and incubated at 37°C for 5 hours. Growth medium was then replaced with fatty acid uptake solution. Kinetic readings were started immediately with a Molecular Devices FlexStation® 3 Reader. A: adipocytes; B: fibroblasts; C: no cells (uptake reagent only).
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Fibroblasts vs. adipocytes; insulin dose-response
3T3 L1 adipocytes were plated at 50,000 cells/well in 100 µl of DMEM/FBS in a 96-well plate and incubated at 37°C for 5 hours, then serum deprived for 1 hour. Different concentrations of insulin were added into the well and incubated for 30 minutes at 37°C, 5% CO2 incubator. At the end of the incubation time, 100 µl of fatty acid mixture was added into the well, and kinetic readings were started immediately with a FlexStation® 3 Reader. Traces A to F correspond to adipocytes with 160, 16, 8, 1.6, 0.16, and 0 nM insulin, respectively; traces G and H correspond to fibroblasts with 0 and 160 nM insulin, respectively.
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Leptin inhibitory effect
3T3L1 adipocytes were plated at 50,000 cells/well/100 µl of DMEM/FBS in 96-well plate for 5 hours, then serum deprived for 1 hour with or without leptin (100 nM). Insulin at 10 nM was then added into the well and incubated for a further 30 minutes at 37°C, 5% CO2 incubator. At the end of the incubation time, 100 µl of fatty acid was added into the well, and kinetic readings were started immediately with a FlexStation® 3 Reader. Data courtesy of Cambridge Biotechnology. A: insulin, B: insulin and leptin, C: basal.
Technology of QBT Fatty Acid Uptake Assay Kit
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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 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.
Resources of QBT Fatty Acid Uptake Assay Kit
eBook
Optimized Assays for Breakthrough Research
Optimized Assays for Breakthrough Research
Biology research has been greatly simplified by the availability of standardized assay kits, instruments, and assay protocols from commercial manufacturers.
Scientific Poster
Quantification of Fatty Acid Uptake in HepG2 Cells and Adipocytes Using the QBT™ Fatty Acid Uptake Assay
Quantification of Fatty Acid Uptake in HepG2 Cells and Adipocytes Using the QBT™ Fatty Acid Uptake Assay
In this poster we demonstrate that QBT Fatty Acid Uptake Assay is well suited for the analysis of LCFA uptake in other mammalian cell types as well as previously shown adipocytes. Fa…
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Scientific Poster
Bringing Primary Cell Relevance to Screening: Clonetics Conditionally Immortalized Cells and QBT™ Fatty Acid Uptake Assay for High Throughput Screening
Bringing Primary Cell Relevance to Screening: Clonetics Conditionally Immortalized Cells and QBT™ Fatty Acid Uptake Assay for High Throughput Screening
This document describes how QBT Fatty Acid Uptake Assay kit provides a single step homogeneous in vitro assay for use in HTS and metabolic disease research. Download to explore more.…
Data Sheet
QBT Fatty Acid Uptake Assay Kit
QBT Fatty Acid Uptake Assay Kit
QBT Fatty Acid Uptake Assay Kit delivers real-time uptake kinetics and is ideally suited for high throughput screening applications. Read this data sheet to explore QBT Fatty Acid ki…
