- High sensitivity and wide linear range (0 – 4 mmol/L)
- A rapid (10 minute) and robust enzyme-based assay which does not require expensive instrumentation
- High reproducibility
The MaxDiscovery™ Non-Esterified Fatty Acids (NEFA) Assay Kit is a plate-based colorimetric enzymatic assay for the quantitative determination of non-esterified fatty acids (NEFA) in serum samples from rodents or other mammals.
The kit uses a spectrophotometric assay to detect non-esterified fatty acids directly from serum samples, enabling researchers to detect non-esterified fatty acids levels in animal serum and other tissue matrices such as urine and ascites.
The MaxDiscovery Non-Esterified Fatty Acids (NEFA) Assay Kit is a simple, accurate and automation-compatible method for measuring NEFA levels in serum samples. The kit is based on the reaction of NEFA in serum with ATP and Coenzyme A (CoA) in the presence of acyl-CoA synthetase (ASC) to form acyl-CoA, AMP and PPi. The acyl-CoA is then oxidized by acyl-CoA oxidase (ACOD) to produce H2O2. The presence of H2O2 and peroxidase (POD) allows 4-aminoantipyrine to react with 3-methyl-N-ethyl-N-(β-hrdroxyenthyl)-aniline (MEHA) to form a purple color. The absorption of this color is measured at 550 nm, and its intensity is directly proportional to the NEFA concentration.
The kit contains sufficient materials to rapidly test 42 serum samples in duplicate, as well as a control solution containing NEFA standard (4 mmol/L) which can be used to calibrate the assay.
Non-esterified fatty acids are the major component of triglycerides, which consist of three fatty acids linked to a glycerol backbone. NEFA is considered as a biomarker of negative energy balance. Low concentrations of NEFA are found in the blood of healthy animals. Increased concentration of NEFA in blood indicates breakdown of lipolysis, which happens when the supply of glucose is insufficient to meet energy needs. Therefore, the measurement of NEFA is essential for reducing the risk of gastrointestinal (displaced abomasum), metabolic (clinical ketosis), and infectious (e.g. metritis) diseases.
Selected Publications that Cite Using the MaxDiscovery Non-esterified Fatty Acid (NEFA) Assay Kit
Baena, M., et al. (2016) Fructose, but not glucose, impairs insulin signaling in the three major insulin-sensitive tissues. Scientific Reports. 6: 26149, doi:10.1038/srep26149.
The MaxDiscovery Non-Esterified Fatty Acid (NEFA) Assay Kit has the capacity for 96 determinations or testing of 42 samples in duplicate (using 12 wells for standards). Store the kit at 4°C. The shelf life is 6 months after receipt when the kit is properly stored.