EnzyChrom™ Free Fatty Acid Assay Kit
Datasheet PDF  MSDS/Bulk Order


Application
For quantitative determination of fatty acid and evaluation of drug effects on its metabolism.

Key Features
Sensitive. Use 10 μL samples. Linear detection range: colorimetric assay 7 - 1000 μM, fluorimetric assay 7 - 100 μM fatty acid.

Convenient. Room temperature "mix-and-read" procedure can be readily automated for high-throughput assay of thousands of samples per day.

Method
OD570nm, or FL530/585nm

Samples
Serum, plasma, urine, saliva, milk, cell cultures, food, agriculture etc

Species
All

Procedure
30 min

Size
100 tests

Detection Limit
7 μM

Shelf Life
6 months

More Details
Fatty acids are aliphatic monocarboxylic acids that are ubiquitously found in animal or vegetable fat, oil and wax. Fatty Acids play important roles in cellular synthesis, energy metabolism and are implicated in diverse disorders such as diabetes mellitus, sudden infant death syndrome and Reye Syndrome. BioAssay Systems’ method provides a simple, one-step and high-throughput assay for measuring free fatty acids. In this assay, free fatty acids are enzymatically converted to acyl-CoA and subsequently to H2O2. The resulting H2O2 reacts with a specific dye to form a pink colored product. The optical density at 570nm or fluorescence intensity (530/585 nm) is directly proportional to the free fatty acid concentration in the sample.

1. Can you tell me what is the conversion factor for free fatty acids from µM to mEq/L? We are about to measure FFA with your assay (EFFA-100) but most publications in domestic animals are in mEq/L?



1 mEq/L = 1 mmole/L = 1000 μM.

2. I have just noticed that EDTA can interfere in this assay but, since I used Microvette CB 300 capillary tubes containing Potassium EDTA to collect the blood, I would like to know at what concentration (if known) EDTA is an inhibitor of the reaction.



EDTA in the sample will complex Mg2+ and Ca2+ ions of the assay buffer, necessary cofactors for the enzymes that are used in the assay.

Plasma EDTA tubes usually contain around 5 mM EDTA in the final volume if filled to the draw volume. Much of the EDTA will be complexed with blood Ca2+/Mg2+ ions, and thus not interfere with the assay. The EDTA concentration will be higher if the tubes are only partially filled.

Since the sample to assay buffer ratio is 1:10 and the assay buffer Mg2+ concentration is 1 mM, I would not expect any interference in samples from tubes that have been filled completely. If the collection tube had only been filled partially, I would recommend supplementing the assay working reagent with an additional 1 mM MgCl2.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
1. Hwang I et al (2012) Catalase deficiency accelerates diabetic renal injury through peroxisomal dysfunction. Diabetes 61(3):728-738. Assay: Fatty Acid in Mouse Plasma (Pubmed).

2. Seo, CW et al (2011). Antihyperlipidemic and body fat-lowering effects of silk proteins with different fibroin/sericin compositions in mice fed with high fat diet. J Agric Food Chem 59(8):4192-4197. Assay: Fatty Acid in Mouse Plasma (Pubmed).

3. Primassin, S., S. Tucci, et al. (2011). Hepatic and muscular effects of different dietary fat content in VLCAD deficient mice. Mol Genet Metab 104(4): 546-51. Assay: Fatty Acid in Mouse Serum (Pubmed).

4. Jelinek D et al (2012) The Niemann-Pick C1 Gene Is Downregulated in Livers of C57BL/6J Mice by Dietary Fatty Acids, but Not Dietary Cholesterol, through Feedback Inhibition of the SREBP Pathway. J Nutr 142(11):1935-1942. Assay: Fatty Acid in Mouse Plasma (Pubmed).

5. Jin Kyubob et al (2012) Dysregulation of hepatic fatty acid metabolism in chronic kidney disease. Nephrol Dial Transplant. Assay: Fatty Acid in Rat Liver (Pubmed).

6. Lu Z et al (2012) TLR4 Antagonist Reduces Early-Stage Atherosclerosis in Diabetic Apolipoprotein E-deficient Mice. J Endocrinol . Assay: Fatty Acid in Mouse Serum (Pubmed).

To find more recent publications, please click here. To find more recent publications, please click here.
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EnzyChrom™ Free Fatty Acid Assay Kit
Catalog No: EFFA-100
Price: $339    Qty:
For orders of 10 or more kits, please contact us at 1-510-782-9988 x 1 for best pricing.

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Shipment: Same Day for order by 2pm PST
Delivery: 1-2 days (US), 3-6 days (Intl);
Storage: -20°C



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