Fatty Acid Oxidation Complete Assay Kit (ab222944)
Key features and details
- Assay type: Cell-based (quantitative)
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Adherent cells, Purified mitochondria, Suspension cells
Overview
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Product name
Fatty Acid Oxidation Complete Assay Kit
See all Fatty Acid Oxidation kits -
Detection method
Fluorescent -
Sample type
Adherent cells, Suspension cells, Purified mitochondria -
Assay type
Cell-based (quantitative) -
Product overview
Fatty Acid Oxidation Complete Assay Kit (ab222944) offers a complete solution to measure Fatty Acid Oxidation (FAO) in live cells using conventional fluorescence plate readers. This product combines Fatty Acid Oxidation Assay (ab217602) and Extracellular Oxygen Consumption Assay (ab197243) in one practical kit for single purchase.
The assay uses the 18C unsaturated fatty acid Oleate as substrate, and includes two FAO modulators, etomoxir and FCCP. Etomoxir, an inhibitor of the carnitine transporter CPT1, prevents Oleate import and thereby limits the supply of reducing equivalents to the ETC, reducing oxygen consumption in turn. The remaining ETC (electron transport chain) activity is driven by non-long chain FAO. FCCP treatment induces maximal ETC activity by dissipating the mitochondrial membrane potential, while the increased demand for reducing equivalents causes a concomitant increase in the FAO activity. If exogenous long-chain fatty acid is unavailable or import is inhibited, FAO activity will be limited.
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Notes
Learn more about the full range of assays to measure glycolysis, oxygen consumption, fatty acid oxidation and metabolic flux in live cells.
Or review the full metabolism assay guide for other assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress.
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Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 55 tests Etomoxir 1 vial Extracellular O2 Consumption Reagent 1 vial FAO Conjugate 1 x 1ml FAO Control 1 x 500µl FAO Tablet 1 tablet FCCP 1 vial High Sensitivity Oil 1 x 15ml L-Carnitine 1 vial -
Research areas
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Alternative names
- Beta Oxidation
- Beta-Oxidation
Images
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FAO-driven oxygen respiration in HepG2 cells.
FAO-driven oxygen respiration in HepG2 cells treated with the CPT-1 inhibitor Etomoxir (white) and uncoupler FCCP (gray).
Untreated cells (basal FAO) curve shows a steady increase of the Extracellular O2 Consumption Reagent signal reflecting ETC-driven oxygen consumption. Signal Control shows probe signal in the absence of cell respiration. Etomoxir treatment prevents oleate import, resulting in reduced availability of reducing equivalents and a resultant decrease in ETC activity. The remaining ETC activity (difference between Etoxomir treatment and Signal Control) is driven by metabolic activity other than long chain FAO. FCCP treatment induces maximal ETC activity by dissipating the mitochondrial membrane potential. Increased demand for reducing equivalents causes a concomitant increase in FAO as indicated by the rapid increase in Extracellular O2 Consumption Reagent signal. This strong increase in ETC activity is not observed where exogenous LCFA is unavailable or where import is inhibited.
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FAO-driven respiration of HepG2 cells treated with etomoxir and FCCP.The figure summarizes the balance between these parameters under “Basal” and “Maximum” (FCCP treated) conditions. The increased energy demand imposed by FCCP treatment is met by increased FAO.
