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Cat.No: CMAK-YJL-0360 Datasheet Instruction for Use
Specification Quantities
Price $961.77–1602.95
| Product Name | Free Fatty Acids Colorimetric Assay Kit |
| Catalog No. | CMAK-YJL-0360 |
| Description | Free fatty acids react with acyl-CoA under the action of acyl-CoA synthetase; acyl-CoA then reacts with acyl oxidase to produce hydrogen peroxide. In the presence of an enzyme and a chromogen, hydrogen peroxide reacts to form a red substance that exhibits maximum absorption at 546 nm; the intensity of the color is directly proportional to the concentration of free fatty acids. This kit is suitable for detecting free fatty acid levels in serum, animal tissues, and cell samples. |
| Kit Components | Reagent 1: Enzyme diluent, 24 mL×1 bottle Reagent 2: Enzyme reagent, powder × 4 vials Reagent 3: Chromogenic agent, 6 mL × 1 bottle Reagent 4: 1 mmol/L Standard, 0.2 mL × 1 vial Microplate: , 96-well × 1 plate |
| Materials Required But Not Provided | Equipment: Microplate reader (540 nm–550 nm; optimal detection wavelength: 546 nm) |
| Reagent Preparation | Before testing, allow the reagents in the kit to equilibrate to room temperature. Preparation of Reagent 2 Working Solution: Take one vial of Reagent 2 and dissolve it in 6 mL of Reagent 1. Incubate at 37°C for 5 minutes before each use. Any unused portion may be stored at 2–8°C, protected from light, for up to 5 days. Incubate Reagent 4 at 37°C for 5 minutes before each use. |
| Sample Preparation | Liquid samples (e.g., serum, plasma): Can be tested directly. Tissue samples: Homogenize according to a ratio of tissue sample weight (g) : volume of saline (0.9% NaCl) (mL) = 1:9. Centrifuge at 4°C and 10,000×g for 10 minutes. Transfer the supernatant to ice for testing, and reserve a portion of the supernatant for protein concentration measurement. Cell samples: Take 3 × 10⁶ cells, add 150 μL of saline (0.9% NaCl), and homogenize. Centrifuge at 4°C, 10,000 × g for 10 minutes. Transfer the supernatant to ice for testing, and reserve a portion of the supernatant for protein concentration determination. Before formal testing, select 2–3 samples expected to show significant differences and dilute them to different concentrations for a preliminary experiment. |
| Test Procedure | Blank wells: Pipette 10 μL of double-distilled water into the blank wells. Standard wells: Pipette 10 μL of 1 mmol/L standard solution into the standard wells. Test wells: Pipette 10 μL of sample into the corresponding test wells. Add 200 μL of Reagent II working solution to the blank, standard, and test wells. Shake the plate on the microplate reader for 10 seconds, then incubate at 37°C for 5 minutes; Add 50 μL of Reagent 3 to each well. Shake the plate on the microplate reader for 10 seconds, then measure the OD value (A1) of each well at 546 nm; Incubate at 37°C for 5 minutes, then measure the OD value (A2) of each well at 546 nm; ΔA = A2 – A1 |
| Storage and Transportation | Store at 2-8°C, protect from light, for 6 months. |
For research use only, not for clinical use.
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