Description
Acetaldehyde Assay Kit (BA0187) (BA0187)
The Acetaldehyde Assay Kit (SKU: BA0187) provides a fast, sensitive fluorimetric 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 generated reduces a probe to make it fluorescent; the fluorescence intensity, measured at 530/585 nm, is directly proportional to the acetaldehyde concentration. With a linear detection range of 0.5 to 60 uM using a 50 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 (BA0187) |
| SKU: | BA0187 |
| Detection Method: | Fluorimetric (lambda ex/em = 530/585 nm) |
| Detection Range: | 0.5 to 60 uM (50 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 sensitive fluorimetric assay for the quantitative determination of acetaldehyde based on aldehyde dehydrogenase catalysed oxidation, in which the NADH generated reduces a probe to a fluorescent product measured at 530/585 nm, proportional to acetaldehyde concentration. The linear detection range is 0.5 to 60 uM (50 uL sample) and the room-temperature assay is read after 30 minutes.
- Fast and sensitive; linear detection range 0.5 to 60 uM (50 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 to process thousands of samples per day
- 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 50 uL of each sample in duplicate into separate wells (one Sample and one Sample Blank). |
| 2 | Prepare Working Reagent (WR) for the four Standards and Sample wells by mixing, per well: 40 uL Assay Buffer, 8 uL NAD Solution, 5 uL Probe, 1 uL Enzyme A and 1 uL Enzyme B. Prepare Blank Working Reagent (BWR) for the Sample Blank wells by mixing, per well: 41 uL Assay Buffer, 8 uL NAD Solution, 5 uL Probe and 1 uL Enzyme B (no Enzyme A). |
| 3 | Prepare standards fresh: mix 5 uL of the 3 M Standard with 995 uL distilled water to make 15 mM acetaldehyde, then mix 4 uL of the 15 mM with 996 uL distilled water to make a 60 uM Premix, and dilute as shown in the table. |
| 4 | Transfer 50 uL standards into separate wells of a black flat-bottom 96-well plate. |
| 5 | Add 50 uL WR to the Standards and Sample wells and 50 uL BWR to the Sample Blank wells. Tap to mix and incubate for 30 minutes at room temperature. |
| 6 | Read fluorescence at 530/585 nm. |
Subtract the blank value (#4) from the standard values and plot delta F against standard concentrations to determine the slope. [Acetaldehyde] = (F_S - F_SB) / Slope x n (uM), where F_S and F_SB are the readings of the Sample and Sample Blank respectively and n is the dilution factor. If the sample fluorescence exceeds that of the 60 uM standard, dilute in water and repeat, multiplying by the dilution factor. Conversions: 1 uM acetaldehyde equals 4.4 ug/L or 44 ppb.
| Component | Quantity | Storage |
| Assay Buffer | 10 mL | -20C |
| Enzyme A | 120 uL | -20C |
| NAD Solution | 1 mL | -20C |
| Enzyme B | 120 uL | -20C |
| Probe | 750 uL | -20C |
| 3 M Standard | 100 uL | -20C to 4C |
