Glucose Assay Kit. Colorimetric / Fluorometric (ab65333/K606)

Glucose Assay Kit ab65333 is a quantitative, addition-only assay with just one 30-min incubation step. In the assay, glucose oxidase produces hydrogen peroxide, with detection via peroxidase. Readout on any colorimetric (570 nm) or fluorometric (Ex/Em = 535/587 nm) plate reader.

- Cited in over 200 publications
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

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Images

Functional Studies - Glucose Assay Kit (AB65333), expandable thumbnail

Publications

Key facts

Detection method: Colorimetric/Fluorometric
Sample types: Urine, Plasma, Cell culture supernatant, Serum, Other biological fluids, Tissue Lysate, Cell Lysate
Assay type: Quantitative
Range: 1 - 10000 µM
Assay time: 40m
Sensitivity: = 1 µM

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Target data

See full target information D-Glucose

What's included?

2000 Test

Components

Assay Buffer 2

20 x 25 mL

Developer Mix B

20 x 1 Vial

Glucose Standard

20 x 100 µL

OxiRed™ Probe

20 x 0.2 mL

Key facts

Detection method: Colorimetric/Fluorometric
Sample types: Urine, Plasma, Cell culture supernatant, Serum, Other biological fluids, Tissue Lysate, Cell Lysate
Assay type: Quantitative
Range: 1 - 10000 µM
Assay time: 40m
Assay Platform: Microplate reader
Sensitivity: = 1 µM

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: -20°C
Appropriate long-term storage conditions: -20°C
Storage information: -20°C

Notes

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 compatible ab102517.

For glucose uptake assays, we provide:

- 2-deoxyglucose-based colorimetric glucose uptake assay Glucose Uptake Assay Kit (Colorimetric) ab136955, and fluorometric glucose uptake assay Glucose Uptake Assay Kit (Fluorometric) 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 2-NBDG Glucose Uptake Assay Kit ab287845 and solid chemical 2-NBDG 2-NBD-Glucose, Fluorescent glucose uptake probe 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 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 compatible 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Associated diseases and disorders

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.

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10 product images

Functional Studies - Glucose Assay Kit (ab65333)
Glucose measured in cell lysates showing quantity (nmol) per million cells.
Samples with the concentration of 2x10⁷ cells/mL were used. Samples were diluted 1.5-13.5 fold and measured colorimetrically.
Image from Shao W et al., PLoS One 7(1), Fig 2C. Doi: 10.1371/journal.pone.0028784. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Functional Studies - Glucose Assay Kit (ab65333)
Shao et al investigated the functional outcome of long- term curcumin supplementation on glucose homeostasis. Glucose metabolism was determined in animals with low fat diet (LFD), high fat diet (HFD) and HFD with curcumin feeding using ab65333. Intraperitoneal insulin tolerance tests(IPITT) were conducted at the end of the 26 weeks. It was concluded curcumin improves insulin sensitivity and disposal of glucose.
Functional Studies - Glucose Assay Kit (ab65333)
Standard curve: mean of duplicates (+/- SD) with background reads subtracted
Functional Studies - Glucose Assay Kit (ab65333)
Standard curve: mean of duplicates (+/- SD) with background reads subtracted
Functional Studies - Glucose Assay Kit (ab65333)
Glucose measured in human biological fluids showing quantity (μmol) per mL of tested sample. Samples were diluted 13.5 fold and measured colorimetrically.
Biochemical assay - Glucose Assay Kit (ab65333)
Diagram showing the principles of the Glucose assay method.
Biochemical assay - Glucose Assay Kit (ab65333)
Shao et al used Lactate assay kit L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330 and Glucose assay kit ab65333 to investigate the impact of Salvigenin, a potential therapeutic) on glycolysis with cell culture supernatant of HepG2 cells.
Salvigenin repressed HCC cell aerobic glycolysis. Salvigenin (25 µM, 50 µM, 100 µM) was utilized to treat HepG2 cells. The level of glucose uptake. The level of lactate production.
The supernatant of the mediums of HepG2 cells was harvested. The floating cells and cell debris were removed through 5 min of centrifugation (1000 g, 4 °C). Then, the supernatant was harvested. As instructed by the manufacturer, the glucose uptake detection kit (ab65333, Abcam, USA) and the lactate production detection kit (L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330, Abcam, USA) were taken to determine the levels of glucose uptake and lactate generation.
Biochemical assay - Glucose Assay Kit (ab65333)
Carreno-Florez et al used Lactate assay kit L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330 and Glucose assay kit ab65333 to investigate glycolytic flux and the Warburg effect in cultured human ΔF508/ΔF508 cystic fibrosis airway epithelial cells (CFBE41o-, hereafter called CF AECs).
Glucose consumption in the basolateral medium of IFN-β-stimulated or influenza virus and respiratory syncytial virus (RSV)-infected CF AECs measured by colorimetric assay. L-lactate concentration measured in apical secretions from CF AECs during IFN-β stimulation or RSV infection measured by colorimetric assay. For all experiments n ≥ 3. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
Glucose consumption (Abcam, ab65333) and lactate secretion (Abcam, L-Lactate Assay Kit (Colorimetric/Fluorometric) ab65330) were measured after 18 h of IFN-β stimulation or 72 h of RSV infection using colorimetric assays. Glucose consumption was determined by subtracting the glucose concentration in basolateral medium from the glucose concentration in the feeding medium. For lactate secretion, MEM without phenol red was added apically for 18 h during IFN-β stimulation or for the last 24 h of RSV infection and was collected to determine the extracellular lactate concentration.
Biochemical assay - Glucose Assay Kit (ab65333)
Glucose colorimetric assay kit ab65333 used to measure glucose in human hepatocellular carcinoma tissue.
Zhou at al measure relative glucose level in 30 paired hepatocellular carcinoma and adjacent nontumoral tissues. The results showed that glucose levels were strikingly decreased in HCC tissues compared to those in adjacent nontumoral tissue samples. Tissues were dissolved in double distilled water and homogenized by ultrasonic treatment. A PCA/KOH deproteinization step was performed before glucose assay. All the steps were conducted as the protocol described.
Biochemical assay - Glucose Assay Kit (ab65333)
Glucose colorimetric assay kit ab65333 used for glucose quantitation in conditioned cell culture media from human cerebral organoids.
Fertan et al established the glucose colorimetric assay kit as a normalization tool for the number of living cells in various cerebral organoid cultures. Image compares remaining glucose levels in cell culture media between low density samples (2 organoids) and high density samples (8-11 organoids). Fertan et al then used normalization using the glucose assay kit as part of measuring small, soluble aggregates of beta-amyloid and tau produced by cerebral organoids in Alzheimer’s disease research.