Free Fatty Acids (NEFA/FFA) Colorimetric Assay Kit (Enzyme Method)
SKU: E-BC-K792-M-96
Free Fatty Acids (NEFA/FFA) Colorimetric Assay Kit (Enzyme Method)
| SKU # | E-BC-K792-M |
| Detection Instrument | Microplate reader (540-550 nm, optimum wavelength: 546 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Detection Principle | Free fatty acids (NEFA/FFA) generate acyl-CoA under the action of acyl synthase, then acyl-CoA generates hydrogen peroxide under the oxidation of acyl oxidase. Hydrogen peroxide reacts in the presence of enzymes and chromogens, producing a red substance with maximum absorption at 546 nm. The amount of this red product is proportional to the content of free fatty acids. |
| Sample Type | Serum, Plasma, Animal tissue, Cell |
| Detection Method | Colorimetric method |
| Detection Instrument | Microplate reader (540-550 nm, optimum wavelength: 546 nm) |
| Research Area | Glycolysis And Lipid Metabolism, Metabolic Diseases, Energy Metabolism |
| Other Reagents Required | Normal saline (0.9% NaCl), Double distilled water |
| Storage | This product can be stored at 2-8°C/-20°C for 12 months with shading light. Please refer to the manual for the specific storage condition of the components. |
| Valid Period | 12 months |
| Sensitivity | 0.14 mmol/L |
| Detection Range | 0.14-4 mmol/L |
| Precision | inter-assay CV: 2.5-6.1% | intra-assay CV: 1.5-4.5% |
| Sample Volume | 10 μL |
| Assay Time | 30 min |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample Type | Dilution Factor |
| Human serum | 1-2 |
| Mouse serum | 4-6 |
| Rat serum | 4-6 |
| Bovine serum | 1-2 |
| 10% Rat liver tissue homogenate | 1-4 |
| 10% Rat heart tissue homogenate | 1-4 |
| 10% Mouse liver tissue homogenate | 1 |
| 3 × 10^6 293T cells | 1 |
The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Enzyme Diluent | 12mL x 1 vial | 24mL x 1 vial | 2-8℃, 12 months |
| Reagent 2 | Enzyme Reagent | Powder x 2 vials | Powder x 4 vials | 2-8℃, 12 months |
| Reagent 3 | Chromogenic Agent | 3mL x 1 vial | 6mL x 1 vial | 2-8℃, 12 months |
| Reagent 4 | 1mmol/L Standard | 0.2mL x 1 vial | 0.2mL x 1 vial | 2-8℃, 12 months |
| Microplate | 96 wells | No requirement | ||
| Plate Sealer | 2 pieces | |||
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.