Free Fatty Acid Assay Kit ab65341 is a no-wash, addition-only assay with two 30 min incubations, used with bodily fluids, culture supernatants, and cell/tissue lysates. Free fatty acids are converted to CoA derivatives, which are oxidized, with colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) readout.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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Publications
- Nature communications 14:69822023Hepatocyte FBXW7-dependent activity of nutrient-sensing nuclear receptors controls systemic energy homeostasis and NASH progression in male mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHui Xia,Catherine R Dufour,Younes Medkour,Charlotte Scholtes,Yonghong Chen,Christina Guluzian,Wafa B'chir,Vincent GiguèrePubMed 37914694
- Journal of biochemical and molecular toxicology 38:e235722023Let-7a-5p abrogates progression of papillary thyroid carcinoma cells by decreasing nuclear receptor subfamily 6 group a member 1-mediated lipogenesis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTao Zhao,Jinghui Sun,Xiangdong Lu,Lingling Liu,Lin Chen,Wei Zhao,Bin ZhouPubMed 37905833
Key facts
- Detection method
Colorimetric/Fluorometric
- Sample types
Cell Lysate, Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Other biological fluids
- Assay type
Quantitative
- Assay time
60m
- Sensitivity
> 2 µM
What's included?
Recommended products
Free Fatty Acid Assay Kit ab65341 is a no-wash, addition-only assay with two 30 min incubations, used with bodily fluids, culture supernatants, and cell/tissue lysates. Free fatty acids are converted to CoA derivatives, which are oxidized, with colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) readout.
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Key facts
- Detection method
Colorimetric/Fluorometric
- Sample types
Cell Lysate, Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum, Other biological fluids
- Assay type
Quantitative
- Assay time
60m
- Assay Platform
Microplate reader
- Sensitivity
> 2 µM
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
- Storage information
-20°C
Notes
Free Fatty Acid Assay Kit ab65341 uses a convenient, sensitive enzyme-based method for detecting long-chain free fatty acids in various mammalian and other samples, such as serum, plasma and other body fluids, food, growth media, etc.
In the free fatty acid assay protocol, fatty acids are converted to their CoA derivatives (coenzyme A), which are subsequently oxidized, leading to formation of color/ fluorescence. Fatty acids can then be easily quantified by either colorimetric (λ= 570 nm) or fluorometric (Ex/Em= 535/587 nm) methods. Palmitic acid is used to generate a standard curve.
Free fatty acid assay protocol summary:
- add samples and standards to wells
- add reaction mix and incubate for 30 min at 37°C
- analyze with microplate reader
Δ Note: Assay time does not include preparation of the samples.
Chinese protocol available. See protocols section below.
Previously called Free Fatty Acid Quantification Assay Kit. This assay detects C-8 (octanoate) and longer fatty acids. Review our to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader. **How other researchers have used Free Fatty Acid Assay Kit ab65341** This Free Fatty Acid assay kit has been used in publications in a variety of sample types, including: - Human: cell culture lysates1, serum2 - Mouse: plasma and liver tissue3, serum4, plasma5, heart tissue6, liver7 - Rat: serum8, plasma9, cytosol of articular chondrocyte primary cell cultures10 - Cell culture medium11 - C *reinhardtii algae12*- Drosophila13 - C elegans14 References: 1 - Ali A et al 2018, Phokrai P et al 2018; 2 - Tahapary DL et al 2018; 3 - Becares et al 2019; 4 - Pan J et al 2019, Maatta J et al 2018, Rohm M et al 2018, Shimazu T et al 2018; 5 - Woo M et al 2018, Tian X et al 2017; 6 - Liu W et al 2017; 7 - Hao et al 2017, Rui W et al 2016; 8 - Honore SM et al 2018; 9 - Van der Werf et al 2018;10 - Lee SW et al 2018; 11 - Kostrzewski T et al 2017; 12 - Ramanan R et al 2018; 13 - Scopelliti A et al 2019; 14 - Pollard AK et al 2019
Supplementary info
- Activity summary
Free Fatty Acids (FFAs) also known as non-esterified fatty acids (NEFAs) are important in various biological processes. These molecules are derived from the hydrolysis of triglycerides and represent an important source of energy. They vary in mass depending on chain length and saturation; common examples include palmitic acid and oleic acid. FFAs exist extensively in plasma as they circulate bound to albumin reflecting their broad expression range across tissues. Researchers measure FFAs using FFA assay kits often employing colorimetric assay techniques in laboratory settings.
- Biological function summary
FFAs play important roles in cellular energy metabolism by serving as substrates for beta-oxidation within mitochondria. They contribute to energy production especially in cardiac and skeletal muscle tissues. FFAs bind to mitochondrial membranes entering beta-oxidation pathways where they undergo sequential enzymatic processing. Although not typically part of large protein complexes FFAs interact with various enzymes and transport proteins facilitating metabolic processes.
- Pathways
FFAs integrate into major metabolic processes such as the citric acid cycle and oxidative phosphorylation pivotal for ATP production. In lipid signalling pathways they serve as ligands for peroxisome proliferator-activated receptors (PPARs) influencing gene expression related to lipid metabolism. PPAR proteins including PPAR-alpha and PPAR-gamma are direct interactors within these pathways modulating lipid homeostasis and insulin sensitivity.
- Associated diseases and disorders
Dysregulation of FFAs links to metabolic syndromes particularly obesity and type 2 diabetes. High levels of plasma FFAs contribute to insulin resistance and are associated with impaired glucose metabolism. In obesity excess FFAs activate inflammatory pathways leading to insulin receptor substrate (IRS) dysfunction. Additionally in type 2 diabetes FFAs impact the function of proteins like AMP-activated protein kinase (AMPK) disrupting glucose uptake and mitochondrial function exacerbating the disease phenotype.
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5 product images
Tian, Xiaoxiang et al., PloS one vol. 12,5 e0176873., Fig 6, doi:10.1371/journal.pone.0176873Functional Studies - Free Fatty Acid Assay Kit - Quantification (ab65341)
Lipolysis were evaluated by measuring free fatty acids released into the culture medium of siCreg1 or si-Scramble 3T3-L1 cells using ab65341, which were treated with BMS-345541 (5 μmol/L), an NF-κB pathway inhibitor, or vehicle (DMSO) for 48 hours prior to lipolysis assessment. ns, no significance; and ***P<0.001.
Samuel P et al., PLoS One 8(1), Fig 7a. DOI: 10.1371/journal.pone.0048425. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Functional Studies - Free Fatty Acid Assay Kit - Quantification (ab65341)
Plasma FFA levels were measured using ab65341 in male and female wild-type (WT) or AT2KO mice either on normal diet (ND) or high fat diet (HFD).
Functional Studies - Free Fatty Acid Assay Kit - Quantification (ab65341)
Func S-1.
Colorimetric standard curve: mean of duplicates (+/-SD) with background readings subtracted.
Functional Studies - Free Fatty Acid Assay Kit - Quantification (ab65341)
Free Fatty Acid measured in biologicals showing concentration (μM).
Functional Studies - Free Fatty Acid Assay Kit - Quantification (ab65341)
Flourometric standard curve: mean of duplicates (+/-SD) with background readings subtracted.
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- Visit troubleshooting
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