Description
Glucose Assay Kit III (BA0243) (BA0243)
The Glucose Assay Kit III (SKU: BA0243) provides a simple, sensitive colorimetric method for determining glucose in a wide range of biological, food and beverage samples. Glucose is a key diagnostic parameter for many metabolic disorders: increased levels are associated with diabetes mellitus and hyperactivity of the thyroid, pituitary and adrenal glands, while decreased levels are found in insulin-secreting tumours, myxoedema, hypopituitarism and hypoadrenalism. This kit uses a single Working Reagent that combines the enzyme reaction and colour reaction in one step, with the colour intensity of the reaction product at 565 nm directly proportional to the glucose concentration. As little as 20 µL of sample is required, and the procedure involves adding a single working reagent and incubating for 30 minutes at room temperature. The assay is well suited to research, drug discovery and food and beverage applications.
| Product Name: | Glucose Assay Kit III (BA0243) |
| SKU: | BA0243 |
| Detection Method: | Colorimetric detection at 565 nm. A single Working Reagent combines the enzyme reaction and colour reaction in one step; the colour intensity of the reaction product is directly proportional to the glucose concentration in the sample. |
| Detection Range: | Linear detection range in a 96-well plate 0.03 to 2 mM (0.54 to 36 mg/dL) glucose |
| Sample Type: | Serum, plasma, urine, saliva, milk, culture medium and other biological samples; also food and beverages |
| Species Reactivity: | All |
| Assay Time: | 30 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | Store all components at -20°C. |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
This assay determines glucose using a single Working Reagent that couples an enzyme reaction with a colour reaction in one step. The reaction product is read at 565 nm, and its colour intensity is directly proportional to the glucose concentration in the sample, quantified against a glucose standard curve.
- Sensitive and accurate. Uses as little as 20 µL sample; linear detection range 0.03 to 2 mM (0.54 to 36 mg/dL) glucose in a 96-well plate.
- Simple and high-throughput. Addition of a single working reagent and a 30-minute incubation at room temperature.
- Direct assays of glucose in serum, plasma, urine, saliva, milk, culture medium and other biological samples.
- Drug discovery and pharmacology: effects of drugs on glucose metabolism.
- Food and beverages: glucose in food, beverages and similar products.
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 | Sample preparation: centrifuge saliva samples for 5 minutes at 14,000 rpm before assay. For milk, mix 100 µL of 6 M HCl with 600 µL milk, centrifuge 5 minutes at 14,000 rpm, transfer the supernatant, add 170 µL of 6 M NaOH per mL supernatant, mix, centrifuge again and assay the supernatant (dilution factor n = 1.36). Samples may be assayed immediately or stored in aliquots at -20°C, avoiding repeated freeze-thaw. |
| 2 | Equilibrate all components to room temperature; keep thawed GDH and Diaphorase refrigerated or on ice during the experiment. |
| 3 | Standards: for the 2 mM standard, mix 48 µL of 300 mg/dL standard with 352 µL dH2O, then dilute to give standards of 2.0, 1.2, 0.6 and 0 mM. Transfer 20 µL standards and samples into separate wells. |
| 4 | Prepare sufficient Working Reagent per standard and sample well by mixing 80 µL Assay Buffer, 1 µL GDH, 1 µL Diaphorase and 8 µL NAD/MTT. Transfer 80 µL Working Reagent into each reaction well and tap the plate to mix briefly and thoroughly. |
| 5 | Incubate 30 minutes at room temperature and read optical density at 565 nm. |
Subtract the blank OD (water, #4) from the standard OD values and plot the change in OD against standard concentrations to determine the slope. [Glucose] = (ODSAMPLE - ODBLANK) / Slope x n mM, where n is the dilution factor. Conversion: 1 mg/dL glucose equals 55.5 µM, 0.001% or 10 ppm. If the calculated concentration exceeds 2 mM, dilute in dH2O, repeat and multiply by the dilution factor.
| Component | Quantity | Storage |
| Assay Buffer | 10 mL | -20°C |
| GDH | 120 µL | -20°C |
| NAD/MTT | 1 mL | -20°C |
| Diaphorase | 120 µL | -20°C |
| Glucose Standard (300 mg/dL) | 1 mL | -20°C |
