Description
D- Mannitol Assay Kit (Colorimetric) (BA0133) (BA0133)
The D-Mannitol Assay Kit (Colorimetric) (SKU: BA0133) provides a simple method for quantifying D-mannitol in food, beverage, agricultural and biological samples. D-Mannitol is a sugar alcohol used as a dietary supplement, sweetener and in intestinal permeability testing for leaky gut, and it also serves as a coating for hard sweets, dried fruits and chewing gums owing to its low ability to attract and hold water. It additionally acts as an osmoprotectant for plants and is used clinically in osmotherapy to reduce intracranial pressure. This assay is based on mannitol dehydrogenase catalysed oxidation of D-mannitol, generating D-fructose and NADH that reduces a formazan (MTT) dye; the intensity of the product colour measured at 565 nm is directly proportional to the D-mannitol concentration. The procedure is a room-temperature, add-mix-read type assay requiring no 37 degrees C heater. The linear detection range is 0.007 to 3 mM mannitol in a 96-well plate assay.
| Product Name: | D- Mannitol Assay Kit (Colorimetric) (BA0133) |
| SKU: | BA0133 |
| Detection Method: | Colorimetric |
| Detection Range: | 0.007 to 3 mM mannitol |
| Sample Type: | Food, beverage, agricultural products, urine and serum |
| 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 is based on mannitol dehydrogenase catalysed oxidation of D-mannitol, which generates D-fructose and NADH that reduces a formazan (MTT) dye. The intensity of the product colour, measured at 565 nm, is directly proportional to the D-mannitol concentration in the sample.
- Fast and sensitive using 20 uL sample, with a linear detection range of 0.007 to 3 mM mannitol in a 96-well plate assay
- Convenient single working reagent procedure with absorbance read after 30 minutes at room temperature; no 37 degrees C heater required
- High-throughput add-mix-read type assay that can be readily automated as a 96-well plate assay for thousands of samples per day
- Direct measurement of mannitol in food, beverage, agricultural products, and biological samples such as urine and serum
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 | Prepare 200 uL 3 mM Premix by mixing 30 uL of the Standard (20 mM) and 170 uL distilled water, then dilute the standards as described in the dilution table. |
| 2 | Transfer 20 uL standards into separate wells of a clear, flat-bottom 96-well plate and transfer 20 uL of each sample into two separate wells, one serving as a sample blank and one as a sample well. |
| 3 | Prepare sufficient Working Reagent for all sample and standard wells by mixing, per well, 85 uL Assay Buffer plus 1 uL Enzyme. |
| 4 | Add 80 uL Working Reagent to the standards and sample wells, and add 80 uL Assay Buffer (no Enzyme) to the sample blank wells, then tap the plate to mix briefly and thoroughly and incubate 30 minutes at room temperature. |
| 5 | Read optical density at 565 nm (520-600 nm). |
Subtract the blank value from the standard values and plot the delta OD against standard concentrations to determine the slope, then calculate [D-Mannitol] = (ODSAMPLE - ODBLANK) / [Slope (mM-1)] x n (mM), where n is the sample dilution factor. Conversions: 1 mM D-mannitol equals 18.2 mg/dL or 182 ppm.
| Component | Quantity | Storage |
| Assay Buffer | 10 mL | -20 degrees C |
| Enzyme | 120 uL | -20 degrees C |
| Standard (20 mM D-Mannitol) | 0.5 mL | -20 degrees C |
