Abcam, ab65341, Free Fatty Acid Assay Kit - Quantification

Size: 100Test / 2000Test
Free Fatty Acid Assay Kit ab65341 is a quantitative, addition-only FFA assay with two 30 min incubations. In the assay, free fatty acids are converted to CoA derivatives, which are oxidized. Readout is on any colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader. - Used with sample types including plasma, serum, cell and tissue extracts, cell culture supernatants, and urine - Cited in over 250 publications. - 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,
Sensitivity:> 2 µM,
Assay time:60m,
Assay Platform:Microplate reader

Product details:
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.
Other names used to refer to this type of assay include NEFA assay and FFA assay.
How the assay works
Free Fatty Acid Assay kit ab65341 measures non-esterified free fatty acids; specifically C-8 (octanoate) and longer fatty acids.
In the free fatty acid assay protocol, free fatty acids are converted to their coenzyme A derivatives by Acyl-CoA Synthetase. These are then oxidized by Acyl CoA Oxidase in a reaction that produces hydrogen peroxide. Hydrogen peroxide is then processed by an enzyme causing a reaction with a probe to generate red color (λmax = 570 nm) and fluorescence (Ex/Em = 535/587 nm). Palmitic acid is used to generate a standard curve.
The free fatty acid assay method using Acyl-CoA Synthetase and Acyl CoA Oxidase as used in ab65341 is the standard method used for free fatty acid assays.
Free fatty acid assay protocol summary
- Add samples and standards to wells.
- Add ACS Reagent and incubate for 30 min at 37°C.
- Add reaction mix and incubate for 30 min at 37°C.
- Analyze with microplate reader.
How other researchers are using 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 lysates
, serum
- Mouse: plasma and liver tissue
, serum
, plasma
, heart tissue
, liver
- Rat: serum
, plasma
, cytosol of articular chondrocyte primary cell cultures
- Cell culture medium
- C reinhardtii algae
- Drosophila
- C elegans
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
Related and recommended products
Free Fatty Acid assay kit ab65341is often used with Triglyceride assay kit
ab65336
, and Cholesterol assay kit
ab65390
, to study lipid metabolism and its role in various metabolic diseases such as diabetes, obesity, liver diseases, and cardiovascular conditions.
Other notes
This product is manufactured by BioVision, an Abcam company, and was previously called K612 Free Fatty Acid Quantification Assay Kit.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.