Description
Glycerol Assay Kit (BA0115) (BA0115)
The Glycerol Assay Kit (SKU: BA0115) provides a simple, direct and automation-ready method for measuring glycerol. Glycerol is widely used in foods, beverages and pharmaceutical formulations and is a major by-product of biodiesel production. The assay uses a single working reagent that combines glycerol kinase, glycerol phosphate oxidase and colour reactions in one step; the colour intensity at 570 nm or fluorescence at 530/585 nm is directly proportional to the glycerol concentration. An optimised formulation gives enhanced reagent and signal stability, and the procedure is compatible with high-throughput screening.
| Product Name: | Glycerol Assay Kit (BA0115) |
| SKU: | BA0115 |
| Detection Method: | Colorimetric (570 nm) / Fluorometric (530/585 nm) |
| Detection Range: | Colorimetric 10 - 1000 uM; fluorometric 2 - 50 uM glycerol |
| Sample Type: | Serum, plasma, food, beverages and pharmaceutical formulations |
| Species Reactivity: | All |
| Assay Time: | 20 minutes |
| Kit Size: | 200 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 12 months after receipt |
| Shipping: | Gel Pack |
A colorimetric or fluorometric assay for glycerol using as little as 10 uL of sample and a single working reagent.
- Sensitive and accurate using as little as 10 uL of sample
- Colorimetric linear range 10 - 1000 uM and fluorometric range 2 - 50 uM glycerol
- Simple single-working-reagent procedure with a 20-minute room-temperature incubation
- Improved reagent and signal stability
- Direct assay of glycerol in biological samples such as serum and plasma
- Studying the effects of drugs on glycerol metabolism
- Glycerol determination in food, beverages and pharmaceutical formulations
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 components to room temperature and keep the thawed Enzyme Mix on ice. Dilute the standard in distilled water per the table (diluted standards can be stored refrigerated for future use). |
| 2 | Transfer 10 uL of each standard and 10 uL of each sample into separate wells of a clear 96-well plate. |
| 3 | For each reaction well mix 100 uL Assay Buffer, 2 uL Enzyme Mix, 1 uL ATP and 1 uL Dye Reagent. Use this Working Reagent on the same day. Transfer 100 uL into each well and tap to mix. |
| 4 | Incubate 20 minutes at room temperature and read optical density at 570 nm (550-585 nm). For the fluorometric assay, use 2 - 50 uM standards and a black plate and read fluorescence at 530/585 nm. |
Subtract the water blank OD from the standard values and plot OD against standard concentration; determine the slope by linear regression. [Glycerol] = (ODSAMPLE - ODH2O) / Slope (mM) for colorimetry, or (F_SAMPLE - F_H2O) / Slope (mM) for fluorimetry. If the sample OD exceeds the 1.0 mM standard, dilute in water, repeat and multiply by the dilution factor. 1 mM glycerol equals 9.2 mg/dL or 92 ppm.
| Component | Quantity | Storage |
| Assay Buffer | 24 mL | -20C |
| Enzyme Mix | 500 uL | -20C |
| ATP | 250 uL | -20C |
| Dye Reagent | 220 uL | -20C |
| Standard (100 mM Glycerol) | 100 uL | -20C |
