Glucose Assay Kit ab65333 is a rapid, simple and sensitive assay used to quantify glucose levels in biological samples such as serum, plasma, and other body fluids, food, growth medium, etc.
How the assay works
In the glucose assay protocol, the glucose enzyme mix oxidizes glucose to generate a product which reacts with a dye to generate color (λ = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The generated color and fluorescence is proportionally to the amount of glucose.
Glucose assay protocol summary
- - Add samples (deproteinized) and standards to wells.
- - Add reaction mix and incubate for 30 min at 37°C.
- - Analyze with microplate reader.
Related Glucose assay products
If you have reducing substances in your samples, we recommend using Glucose Assay Kit - reducing agent compatible Glucose Assay Kit - reducing agent compatibleab102517.
For glucose uptake assays, we provide:
Related and recommended products
Review our metabolism assays overview to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress here.
Glucose assay methods
There are 3 main enzymatic glucose assay methods, based on different enzymes that act on glucose.
- a) Glucose oxidase-based assays, such as ab65333, that rely on the production of hydrogen peroxide by glucose oxidase. This is then followed by the oxidation of a substrate by a peroxidase using the hydrogen peroxide to produce a colorimetric or fluorometric readout. This method is also referred to as the GOD-POD glucose assay method (GOD-POD = glucose oxidase-peroxidase).
- b) Glucose dehydrogenase-based assays, such as Glucose Assay Kit - reducing agent compatibleab102517, that rely on the production of NADH from NAD as part of the action of glucose dehydrogenase on glucose. The increase in NADH can be measured by absorbance at 340nm, or indirectly using the reduction of a tetrazolium dye by NADH to produce a colored or fluorescent product.
- c) Hexokinase-based assays, rely on the production of glucose-6-phosphate by hexokinase from glucose and ATP. Glucose-6-phosphate dehydrogenase then acts on glucose-6-phosphate and NAD or NADP to produce NADH or NADPH, which can be measured in the same way as in glucose dehydrogenase-based assays.
Older glucose assay methods, that are now less commonly used, include:
- - Reducing methods, relying on the ability of glucose to reduce a metal ion when glucose is oxidized. This method is non-specific and any strong reducing agent present in the sample will result in an increased signal.
- - Condensation with o-toluidine, where the aldehyde group of glucose reacts with o-toluidine to form a glucosamine with a green color. Mannose and galactose tend to cross-react with o-toluidine, however these are found in limited quantities in many sample types. The major disadvantage of this assay is that o-toluidine is corrosive and toxic.
Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K606 Glucose Assay Kit.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
