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

Data of QBT Fatty Acid Uptake Assay Kit

  • Fibroblast vs. adipocytes

    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).

  • 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.

    Fibroblasts vs. adipocytes; insulin dose-response

  • Leptin inhibitory effect

    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

  • QBT Fatty Acid Uptake Assay Principle

    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.

Ordering Options of QBT Fatty Acid Uptake Assay Kit

QBT Fatty Acid Uptake Assay Kit

Evaluation Kit

  • 1 Kit = 1 plate x 2 vials each

#R6132

$164.00 USD

1

Explorer Kit

  • 1 Kit = 1 plate x 10 vials

#R8132

$1,094.00 USD

1

Bulk Kit

  • 1 Kit = 5 plates x 10 vials

 

Compatible with 96- and 384-well plate formats.

#R8133

$2,553.00 USD

1

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We are ready to help you solve your tough research challenges. Our proven solutions and highly qualified teams across the globe can help advance your next big discovery.