Description
Acetaldehyde Assay Kit (BA0185) (BA0185)
The Acetaldehyde Assay Kit (SKU: BA0185) provides a fast, sensitive colorimetric method for the quantitative determination of acetaldehyde in biological and food or beverage samples. Acetaldehyde is one of the most widely occurring aldehydes in nature and a metabolic by-product of ethanol in the liver; its accumulation, associated with deficiency of aldehyde dehydrogenase, is linked to facial and body flushing and the effects of hangovers, and it is also naturally present in foods such as ripe fruit, coffee and wine. The assay is based on aldehyde dehydrogenase catalysed oxidation of acetaldehyde, in which the NADH formed reduces a formazan reagent; the intensity of the product colour, measured at 565 nm, is directly proportional to the acetaldehyde concentration. With a linear detection range of 2 uM to 2 mM using a 20 uL sample and a simple single-working-reagent room-temperature procedure read after 30 minutes, the homogeneous format can be readily automated. It is intended for research use only.
| Product Name: | Acetaldehyde Assay Kit (BA0185) |
| SKU: | BA0185 |
| Detection Method: | Colorimetric (OD565 nm) |
| Detection Range: | 2 uM to 2 mM acetaldehyde in 96-well plate assay (20 uL sample) |
| Sample Type: | Plasma, serum, urine, tissue, culture media, and food/beverage samples such as wine, coffee and juice |
| Species Reactivity: | All |
| Assay Time: | 30 min |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
A colorimetric assay for the quantitative determination of acetaldehyde based on aldehyde dehydrogenase catalysed oxidation, in which the NADH formed reduces a formazan reagent to yield a coloured product measured at 565 nm proportional to acetaldehyde concentration. The linear detection range is 2 uM to 2 mM (20 uL sample) and the room-temperature assay is read after 30 minutes.
- Fast and sensitive; linear detection range 2 uM to 2 mM acetaldehyde (20 uL sample)
- Convenient single-working-reagent procedure read after 30 minutes at room temperature
- No 37C heater is needed
- Homogeneous mix-incubate-measure format readily automated on high-throughput systems
- Determination of acetaldehyde in biological samples such as plasma, serum, urine, tissue and culture media
- Determination of acetaldehyde in food and beverage samples such as wine, coffee and juice
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 | Transfer 20 uL of each sample in duplicate into separate wells of a clear flat-bottom 96-well plate (one Sample well and one Sample Blank well). |
| 2 | Prepare Working Reagent (WR) for the four Standards and Sample wells by mixing, per well: 75 uL Assay Buffer, 8 uL NAD/MTT, 1 uL Enzyme A and 1 uL Enzyme B. Prepare Blank Working Reagent (BWR) for the Sample Blank wells by mixing, per well: 75 uL Assay Buffer, 8 uL NAD/MTT and 1 uL Enzyme B (no Enzyme A). |
| 3 | Prepare standards fresh: mix 5 uL of the 3 M Standard with 495 uL distilled water to make 30 mM acetaldehyde, then mix 20 uL of the 30 mM with 280 uL distilled water to make a 2 mM Premix, and dilute as shown in the table. |
| 4 | Transfer 20 uL standards into separate wells. |
| 5 | Add 80 uL WR to the Standards and Sample wells and 80 uL BWR to the Sample Blank wells. Tap the plate to mix and incubate for 30 minutes at room temperature. |
| 6 | Read optical density at 565 nm (520-600 nm). |
Subtract the blank value (#4) from the standard values and plot delta OD against standard concentrations to determine the slope. [Acetaldehyde] = (ODS - ODSB) / Slope x n (mM), where ODS and ODSB are the readings of the Sample and Sample Blank respectively and n is the dilution factor. If the sample OD exceeds that of the 2 mM standard, dilute in water and repeat, multiplying by the dilution factor. Conversions: 1 mM acetaldehyde equals 4.4 mg/dL or 44 ppm.
| Component | Quantity | Storage |
| Assay Buffer | 10 mL | -20C |
| Enzyme A | 120 uL | -20C |
| NAD/MTT Solution | 1 mL | -20C |
| Enzyme B | 120 uL | -20C |
| 3 M Standard | 100 uL | -20C |
