Metabolism Assays
Glucose Oxidase Assay Kit (BA0116)
- SKU:
- BA0116
- Product Type:
- Assay
- Instrument:
- Microplate Reader
- Sample Type:
- Cell, Tissue Lysate, Cell Culture Media
- Research Area:
- Diabetes & Obesity
- Research Area:
- Plant & Environmental Stress
- Research Area:
- Enzyme Activity
Description
Glucose Oxidase Assay Kit (BA0116) (BA0116)
The Glucose Oxidase Assay Kit (SKU: BA0116) provides a simple, direct and high-throughput method for measuring glucose oxidase activity. Glucose oxidase catalyses the oxidation of D-glucose to D-glucono-delta-lactone and is widely used in electrochemical glucose sensors. The assay uses a single working reagent that combines the glucose oxidase reaction with a colour reaction in one step; the change in colour intensity at 570 nm or fluorescence at 530/585 nm is directly proportional to the glucose oxidase activity in the sample. The room-temperature procedure suits both colorimetric and fluorometric formats.
| Product Name: | Glucose Oxidase Assay Kit (BA0116) |
| SKU: | BA0116 |
| Detection Method: | Colorimetric (570 nm) / Fluorometric (530/585 nm) |
| Detection Range: | Colorimetric 0.02 - 10 U/L; fluorometric 0.002 - 1.5 U/L (20 min at 25C) |
| Sample Type: | Cell lysate, culture medium and other biological samples |
| Species Reactivity: | All |
| Assay Time: | 20 minutes |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
A colorimetric or fluorometric kinetic assay for glucose oxidase activity using an H2O2 standard curve and a single working reagent.
- Sensitive and accurate using as little as 20 uL of sample
- Colorimetric range 0.02 - 10 U/L and fluorometric range 0.002 - 1.5 U/L for a 20-minute incubation at 25C
- Simple, high-throughput single-working-reagent procedure
- Direct assay of glucose oxidase activity in cell lysate, culture medium and other biological samples
- Studying the effects of drugs on glucose metabolism
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 | Samples can be analysed immediately or stored in aliquots at -20C; avoid repeated freeze-thaw and centrifuge if particulates are present. Equilibrate components to room temperature and keep the thawed Enzyme on ice. |
| 2 | H2O2 Standard Curve: mix 5 uL 3% H2O2 with 914 uL distilled water, then 20 uL of this with 460 uL distilled water to give 200 uM H2O2. Prepare standards per the table. |
| 3 | Transfer 20 uL of each standard and sample into separate wells. |
| 4 | Working Reagent: mix per well 75 uL Assay Buffer, 10 uL 2 M Glucose, 1 uL HRP Enzyme and 1 uL Dye Reagent. Transfer 80 uL into each well and tap to mix. |
| 5 | Read optical density immediately (OD0) at 570 nm (550-585 nm), incubate 20 minutes at room temperature and read again (OD20). For the fluorometric assay, use a black plate and standards at 20, 12, 6 and 0 uM H2O2 and read fluorescence at 530/585 nm. |
Subtract the water blank OD20 or F20 from the standard values and plot delta-OD or delta-F against concentration to obtain the slope. Compute delta-R_Sample (OD20-OD0 or F20-F0) and delta-R_Blank, then calculate activity using these values, the standard curve slope, the incubation time (20 min) and the dilution factor n. One U/L catalyses 1 umole of H2O2 per minute at pH 7.0 and room temperature. If activity exceeds the range, dilute and repeat; for low activity, extend the incubation.
| Component | Quantity | Storage |
| Assay Buffer | 10 mL | -20C |
| Glucose (2 M) | 1.5 mL | -20C |
| HRP Enzyme | 120 uL | -20C |
| Dye Reagent | 120 uL | -20C |
| Standard (3% H2O2) | 100 uL | -20C |
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