Description
Acetate Assay Kit (Colorimetric and Fluorometric) (BA0140) (BA0140)
The Acetate Assay Kit (Colorimetric and Fluorometric) (SKU: BA0140) provides a straightforward enzyme-coupled method for measuring acetate in biological, food, agricultural and environmental samples. Acetate is a common anion fundamental to all forms of life and, when bound to coenzyme A, is central to the metabolism of carbohydrates and fats. Its acid form, acetic acid, is produced by acetic acid bacteria found universally in foodstuffs, water and soil, and is the main component of vinegar as well as being extensively used in food, dyes, paints, glue and synthetic fibres. This assay uses enzyme-coupled reactions to form a coloured, fluorescent product, with the colour absorbance at 570 nm or fluorescence intensity at 530/585 nm being directly proportional to the acetate concentration in the sample. The detection range is 0.20 to 20 mM acetate for colorimetric assays and 0.13 to 2 mM for fluorometric assays.
| Product Name: | Acetate Assay Kit (Colorimetric and Fluorometric) (BA0140) |
| SKU: | BA0140 |
| Detection Method: | Colorimetric and Fluorometric |
| Detection Range: | 0.20 to 20 mM (colorimetric); 0.13 to 2 mM (fluorometric) |
| Sample Type: | Serum/plasma, food, agriculture and environmental samples |
| Species Reactivity: | All |
| Assay Time: | 30 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20 degrees C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
This kit uses enzyme-coupled reactions to form a coloured, fluorescent product. The colour absorbance at 570 nm or fluorescence intensity at 530/585 nm is directly proportional to the acetate concentration in the sample.
- Uses as little as 10 uL sample with a detection range of 0.20 to 20 mM acetate for colorimetric assays and 0.13 to 2 mM for fluorometric assays
- Provides both colorimetric and fluorometric detection formats
- Enzyme-coupled reactions form a coloured, fluorescent product for straightforward quantitation
- Direct measurement of acetate in biological samples such as serum/plasma, and in food, agriculture and environmental samples
- Drug discovery and pharmacology studies of the effects of drugs on acetate metabolism
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Equilibrate all components to room temperature, briefly centrifuge tubes, and reconstitute Enzyme A with 600 uL Developer and Enzyme B with 120 uL Assay Buffer, ensuring both are fully dissolved and kept refrigerated or on ice during use. |
| 2 | Prepare 400 uL 2 mM Standard by mixing 4 uL 200 mM standard with 396 uL distilled water, then dilute the standard as described in the dilution table. |
| 3 | Transfer 10 uL standards and 10 uL samples into separate wells of a black flat-bottom 96-well plate (or a clear flat-bottom plate for the colorimetric procedure). |
| 4 | Prepare Working Reagent for each reaction well by mixing 90 uL Assay Buffer, 5 uL Enzyme A, 1 uL Enzyme B, 1 uL Dye Reagent and 1 uL ATP, using it within 20 minutes, then transfer 90 uL to each well, mix immediately and incubate for 30 minutes at room temperature. |
| 5 | Read fluorescence intensity at 530/585 nm, or for the colorimetric procedure read OD at 570 nm (550-585 nm). |
Subtract the water blank (Std #5) value from all the standard and sample values, plot the delta F or delta OD of the standards against the standard concentrations, and determine the acetate concentration of samples from the standard curve. Conversions: 1 mM acetate equals 5.9 mg/dL, 0.0059% or 59 ppm.
| Component | Quantity | Storage |
| Assay Buffer | 25 mL | -20 degrees C |
| Developer | 1 mL | -20 degrees C |
| Dye Reagent | 120 uL | -20 degrees C |
| ATP | 120 uL | -20 degrees C |
| Enzyme A (Dried) | 1 tube | -20 degrees C |
| Enzyme B (Dried) | 1 tube | -20 degrees C |
| Standard | 1 mL | -20 degrees C |
