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Non-esterified Free Fatty Acids (NEFA) Assay Kit

Catalog number:E-BC-K014
Pack Size: 96 Tests
Detection method: Colorimetric method 
Detection instrument: Microplate reader, Biochemistry analyzer 
Valid period: 6 months

Non-esterified Free Fatty Acids (NEFA) Colorimetric Assay Kit can be used for detection of non-esterified free fatty acids (NEFA) content in serum, plasma, tissue homogenate, cells or cell supernatant samples.

Detection principle

NEFA and can react with coenzyme A and form acetyl-CoA under the catalysis of acetyl-CoA-synthetase (ACS). Acetyl-CoA can produce H2O2 when catalyzed by acetyl-CoA-oxidase (ACOD). Then H2O2 react with TOOS and 4-amino-antipyrine (4-APP) to generate a colored substrate under the catalysis of peroxidase (POD). The colored substrate has a maximum absorption peak at 546 nm. Measure the OD value at 546 nm and calculate the NEFA content indirectly.


Experimental instrument

Semi-automatic biochemical analyzer (546 nm), automatic biochemical analyzer (546 nm), Microplate reader (546 nm).


Storage and valid period

This kit can be stored at 2~8in the dark for 6 months. Prevent the reagents from freezing.

Please store the opened reagents can be stored for 2 weeks at 2~8in the dark.


Sample treatment

1.    Serum or plasma:

 Separate serum or plasma just in time after blood collection and avoid of hemolysis. It is recommended to detect the sample immediately. (The concentration of NEFA may           increase due to the degradation of lipid.)

2.   Tissue sample:

 Mince the tissues to small pieces, then weighed and homogenized in normal saline on ice, the volume of normal saline (mL): the weight of the tissue (g) =9:1. The tissue            homogenate is centrifuged at 2500 rpm for 10 min and take the supernatant for detection. Meanwhile, determine the protein concentration of supernatant (E-BC-K318, E-BC-   K168, E-BC-K165).

3.   Cell sample:

      Collect cells and treat the sample with mechanical homogenate or sonication on ice. Prepared cell homogenate does not require centrifugation. Meanwhile, determine the    protein concentration of supernatant (E-BC-K318, E-BC-K168, E-BC-K165).


4.   Cell culture supernatant:

 Detect directly.

[Note]: Samples (serum, plasma) can be stored at 2~8for 3 days. It is recommended that the samples should be stored at -20? or lower temperature condition if can’t detect immediately. Tissue homogenate and cell homogenate must be detected in that very day.Don’t use plasma sample anticoagulated with heparin.


Note: The product is for lab research use only (RUO). Not for Human Use. 


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