Description
Glucose Assay Kit II (BA0242) (BA0242)
The Glucose Assay Kit II (SKU: BA0242) is a simple, direct and high-throughput colorimetric assay for the quantitative determination of glucose in a wide range of biological and food samples. Glucose (C6H12O6) 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. The kit uses an enzyme to reduce NAD, and the resulting NADH, measured at 340 nm, is proportional to the glucose concentration in the sample. The room-temperature 'mix-incubate-measure' procedure requires only a single working reagent and a 20-minute incubation, and can be automated on HTS liquid-handling systems for thousands of samples per day.
| Product Name: | Glucose Assay Kit II (BA0242) |
| SKU: | BA0242 |
| Detection Method: | Colorimetric (OD340 nm) |
| Detection Range: | 0.1 to 3 mM (1.8 to 54 mg/dL) glucose (96-well plate) |
| Sample Type: | ['Serum', 'Plasma', 'Urine', 'Saliva', 'Milk', 'Culture medium', 'Food', 'Beverages'] |
| Species Reactivity: | All |
| Assay Time: | 20 min |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20°C |
| Shelf Life: | 6 months |
| Shipping: | Gel Pack |
A colorimetric assay kit for the quantitative determination of glucose in serum, plasma, urine, saliva, milk, culture medium, food and beverages. The kit uses an enzyme to reduce NAD; the produced NADH, measured at 340 nm, is proportional to the glucose concentration, with a linear detection range of 0.1 to 3 mM in a 96-well plate.
- Sensitive and accurate. Use as little as 20 µL samples. Linear detection range in a 96-well plate: 0.1 to 3 mM (1.8 to 54 mg/dL) glucose for colorimetric assays.
- Convenient. Room temperature assay; no 37°C heater is needed.
- Simple and high-throughput. Involves addition of a single working reagent and a 20-min incubation; can be readily automated on HTS liquid-handling systems for thousands of samples per day.
- Direct assays of glucose in serum, plasma, urine, saliva, milk, culture medium and other biological samples.
- Drug discovery/pharmacology: effects of drugs on glucose metabolism.
- Food and beverages: glucose in food, beverages etc.
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 all components to room temperature. During the experiment, keep thawed GDH in a refrigerator or on ice. |
| 2 | Standards and samples: for the 3 mM standard, mix 72 µL 300 mg/dL standard with 328 µL dH2O, then dilute in dH2O to give 3.0, 1.8, 0.9 and 0 mM points (see Standard Dilution table). Transfer 20 µL standards and samples into separate wells. |
| 3 | Prepare sufficient Working Reagent by mixing, per well, 80 µL Assay Buffer, 1 µL GDH and 8 µL NAD in a clean tube. Transfer 80 µL Working Reagent into each reaction well and tap the plate briefly and thoroughly to mix. |
| 4 | Incubate 20 min at room temperature. Read optical density at 340 nm. |
Subtract the blank OD (water, #4) from the standard OD values and plot ΔOD against standard concentrations. Determine the slope and calculate: [Glucose] = (ODSAMPLE – ODBLANK) / Slope × n (mM), where ODSAMPLE and ODBLANK are the optical density values of the sample and water respectively and n is the sample dilution factor. Conversions: 1 mg/dL glucose equals 55.5 µM, 0.001% or 10 ppm. If the calculated glucose concentration exceeds 3 mM, dilute the sample in water and repeat, multiplying the result by the dilution factor.
| Component | Quantity | Storage |
| Assay Buffer | 10 mL | -20°C |
| GDH | 120 µL | -20°C |
| NAD | 1 mL | -20°C |
| Glucose Standard (300 mg/dL) | 1 mL | -20°C |
