Description
Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Urine, Plasma, Cell culture supernatant, Serum, Other biological fluids, Tissue Lysate, Cell Lysate,
Assay type:Quantitative,
Sensitivity:= 1 µM,
Range:1 - 10000 µM,
Assay time:40m,
Assay Platform:Microplate reader
Product details:
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
ab102517
For glucose uptake assays, we provide:
- 2-deoxyglucose-based colorimetric glucose uptake assay
ab136955
, and fluorometric glucose uptake assay
ab136956
. 2-deoxyglucose is metabolized to 2-DG-6-phosphate, which accumulates within cells and can be quantified with an enzymatic assay.
- We also provide 2-NBDG Glucose Uptake Assay Kit
ab287845
and solid chemical 2-NBDG
ab146200
. 2-NBDG is a fluorescent glucose analog that accumulates in cells.
Related and recommended products
Review our metabolism assays overview to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress
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
ab102517
, 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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Glucose often referred to as blood sugar is a simple sugar and an essential carbohydrate in biology. It has a molecular mass of 180.16 g/mol and is highly soluble in water. Glucose circulates in the bloodstream and is absorbed by tissues mainly liver muscle and adipose tissue. It serves as a critical energy source and cells use glucose uptake processes to transport glucose across their membranes. Various diagnostic tools and kits such as glucose assay kits and glucose test kits help measure glucose levels in biological samples.
Biological function summary
Glucose serves as the primary energy substrate for cells providing energy through glycolysis and oxidative phosphorylation. It is not part of any protein complexes but it interacts with numerous enzymes and proteins to regulate metabolic processes. Glucose operates in maintaining homeostasis and the brain relies on it almost exclusively for energy. Glucose assay reagents and glucose detection kits are utilized to quantify glucose concentrations in research studies examining these functions.
Pathways
Glucose is a central component in glycolysis and the tricarboxylic acid (TCA) cycle. In glycolysis glucose is broken down into pyruvate generating ATP and NADH in the process. This pathway involves key regulatory proteins such as hexokinase and phosphofructokinase. In the TCA cycle glucose metabolites further produce ATP and CO2 involving enzymes like citrate synthase. Glucose uptake assays provide insights into how these pathways operate under various physiological conditions.
Glucose regulation and metabolism are tightly linked to diabetes mellitus and metabolic syndrome. Diabetes mellitus is characterized by impaired glucose uptake and insulin regulation often involving insulin receptor pathways. Persistent high glucose levels lead to complications such as neuropathy and retinopathy. AMP-activated protein kinase (AMPK) plays an important role in metabolic syndrome by affecting glucose uptake and energy balance. Understanding glucose's role in these diseases is central to devising therapeutic strategies and interventions.
