Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric/Fluorometric
- Platform: Microplate reader
- Assay time: 60 min
- Sample type: Cell culture supernatant, Cell Lysate, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
- Sensitivity: 2 µM
Product nameFree Fatty Acid Assay Kit - Quantification
See all Free Fatty Acid kits
Sample typeCell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate
Sensitivity> 2 µM
Assay time0h 60m
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 37ºC
- analyze with microplate reader
Δ Note: Assay time does not include preparation of the samples.
Chinese protocol available. See protocols section below.
This product is manufactured by BioVision, an Abcam company and was previously called K612 Free Fatty Acid Quantification Colorimetric/Fluorometric Kit. K612-100 is the same size as the 100 test size of ab65341.
This assay detects C-8 (octanoate) and longer fatty acids.
Review our Metabolism Assay Guide 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
- 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
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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 100 tests 2000 tests ACS Reagent Blue 1 vial 20 vials Enhancer Purple 1 x 200µl 20 x 200µl Fatty Acid Assay Buffer WM 1 x 25ml 20 x 25ml Fatty Acid enzyme mix Green 1 vial 20 vials Fatty Acid Probe (in DMSO) Red 1 x 200µl 20 x 200µl Palmitic Acid Standard (1nmol/µl) Yellow 1 x 300µl 20 x 300µl
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Fatty acids
RelevanceFatty acids are carboxylic acids with long hydrocarbon chains. The hydrocarbon chain length may vary from 8-30 carbons. Fatty acids play very important roles in normal metabolism and many disease development. They are precursors to a number of bioactive classes of compounds such as prostaglandins, leucotrienes and others, and have been implicated in diverse functions such as autism, immune system and inflammation response.
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.
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).
Colorimetric standard curve: mean of duplicates (+/-SD) with background readings subtracted.
Free Fatty Acid measured in biologicals showing concentration (µM).
Flourometric standard curve: mean of duplicates (+/-SD) with background readings subtracted.
ab65341 has been referenced in 227 publications.
- Maier G et al. Transcriptomic, proteomic and phosphoproteomic underpinnings of daily exercise performance and zeitgeber activity of training in mouse muscle. J Physiol 600:769-796 (2022). PubMed: 34142717
- Bhat N et al. Dyrk1b promotes hepatic lipogenesis by bypassing canonical insulin signaling and directly activating mTORC2 in mice. J Clin Invest 132:N/A (2022). PubMed: 34855620
- Pham DV & Park PH Adiponectin triggers breast cancer cell death via fatty acid metabolic reprogramming. J Exp Clin Cancer Res 41:9 (2022). PubMed: 34986886
- Xu Z et al. Intermittent Fasting Improves High-Fat Diet-Induced Obesity Cardiomyopathy via Alleviating Lipid Deposition and Apoptosis and Decreasing m6A Methylation in the Heart. Nutrients 14:N/A (2022). PubMed: 35057432
- Dai B et al. Macrophages in epididymal adipose tissue secrete osteopontin to regulate bone homeostasis. Nat Commun 13:427 (2022). PubMed: 35058428