Glucose Assay Kit

Catalog No. ABIN1000263

Product Details

Target: Glucose

Application: Biochemical Assay (BCA)

Sample Type: Serum, Plasma, Food, Beverages

Specificity: 0.7 mg/dL (39 μM)

Characteristics: Sensitive and accurate. Use as little as 5 µL samples. Linear detection range 0.7 mg/dL (39 µM) to 300 mg/dL (16.6 mM) glucose in 96-well plate.
Simple and convenient. The procedure involves addition of a single working reagent and incubation for 8 min in a boiling water bath.
Improved reagent stability. The optimized formulation has greatly enhanced the reagent and signal stability.
Low interference in biological samples.
No pretreatments are needed. Assays can be directly performed on serum and plasma samples.

Components: Reagent: 50 ml. Standard: 1 ml 300 mg/dL.

Material not included: Pipeting devices, centrifuge tubes, boiling water bath, tube holder. Clear bottom 96-well plates (e.g. Corning Costar) and plate reader. Spectrophotometer and Cuvets for measuring OD at 620-650nm.

Application Details

Application Notes: Direct Assays: glucose in biological samples (e.g. serum and plasma).
Drug Discovery/Pharmacology: effects of drugs on glucose metabolism.
Food and Beverages: glucose in food, beverages etc.

Protocol: THE REAGENT CONTAINS ACETIC ACID. THIS ASSAY IS PREFERABLY CARRIED OUT IN A CHEMICAL FUME HOOD.

Procedure using 96-well plate:
1. Dilute standard in distilled water. Set up1.5-mL centrifuge tubes. Transfer 5 µL diluted standards and samples to appropriately labeled tubes. Transfer 500 µL Reagent to each tube. Close the tubes tightly and mix. Store diluted standards at -20°C for future use.
2. Place the tubes in a tube holder and heat in a boiling water bath or heat block for 8 min. Cool down in cold water bath for 4 min.
3. Transfer 200 µL in duplicate into a clear bottom 96-well plate. Careful: avoid bubble formation. Read optical density at 620-650nm (peak absorbance at 630nm).

Procedure using cuvette:
1. Dilute standards and transfer 12 µL water blank, Standards and samples to appropriately labeled tubes. Transfer 1200 µL Reagent to each tube. Close the tubes tightly and mix.
2. Place the tubes in a tube holder and heat in a boiling water bath for 8 min. Cool down in cold- water bath for 4 min.
3. Transfer 1000 µL reaction mixture into cuvet. Read optical density at 620-650nm (peak absorbance at 630nm) against blank. Note:
1. if the Sample OD is higher than the Standard OD at 300mg/dL, dilute sample in water and repeat the assay.
2. To determine low glucose concentrations, use 50 µL sample and standards (instead of 5 µL) per 500 µL Reagent.

Calculation of Results:

Subtract blank OD (water, #5) from the standard OD values and plot the OD against standard concentrations. Determine the slope using linear regression fitting. Typical serum/plasma glucose values: 70 - 110 mg/dL.
Conversions: 1mg/dL glucose equals 55.5 µM, 0.001% or 10 ppm.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Hedbacker, Birsoy, Wysocki, Asilmaz, Ahima, Farooqi, Friedman: "Antidiabetic effects of IGFBP2, a leptin-regulated gene." in: Cell metabolism, Vol. 11, Issue 1, pp. 11-22, (2010) (PubMed).

Vaitheesvaran, LeRoith, Kurland: "MKR mice have increased dynamic glucose disposal despite metabolic inflexibility, and hepatic and peripheral insulin insensitivity." in: Diabetologia, Vol. 53, Issue 10, pp. 2224-32, (2010) (PubMed).

Olsen, Sarras, Intine: "Limb regeneration is impaired in an adult zebrafish model of diabetes mellitus." in: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society, Vol. 18, Issue 5, pp. 532-42, (2010) (PubMed).

Li, Xie, Qin, Jeha, Saha, Yan, Haueter, Chan, Tsai, Tsai: "The nuclear orphan receptor COUP-TFII plays an essential role in adipogenesis, glucose homeostasis, and energy metabolism." in: Cell metabolism, Vol. 9, Issue 1, pp. 77-87, (2009) (PubMed).

Sekowska, Masson, Celani, Danchin, Vergassola: "Repulsion and metabolic switches in the collective behavior of bacterial colonies." in: Biophysical journal, Vol. 97, Issue 3, pp. 688-98, (2009) (PubMed).

Chiu, Radisic: "Scaffolds with covalently immobilized VEGF and Angiopoietin-1 for vascularization of engineered tissues." in: Biomaterials, Vol. 31, Issue 2, pp. 226-41, (2009) (PubMed).

Cirrik, Oner: "The effect of heavy muscle activity on renal cytoresistance in rats." in: Renal failure, Vol. 31, Issue 8, pp. 683-9, (2009) (PubMed).

Zheng, Pan, Zhang, Labavitch, Wang, Teter, Jenkins: "Evaluation of different biomass materials as feedstock for fermentable sugar production." in: Applied biochemistry and biotechnology, Vol. 137-140, Issue 1-12, pp. 423-35, (2008) (PubMed).

Sullivan, Harris, Palmer: "Convection and hemoglobin-based oxygen carrier enhanced oxygen transport in a hepatic hollow fiber bioreactor." in: Artificial cells, blood substitutes, and immobilization biotechnology, Vol. 36, Issue 4, pp. 386-402, (2008) (PubMed).

Stites, Storms, Bauerly, Mah, Harris, Fascetti, Rogers, Tchaparian, Satre, Rucker: "Pyrroloquinoline quinone modulates mitochondrial quantity and function in mice." in: The Journal of nutrition, Vol. 136, Issue 2, pp. 390-6, (2006) (PubMed).

Jatana, Giri, Ansari, Elango, Singh, Singh, Khan: "Inhibition of NF-kappaB activation by 5-lipoxygenase inhibitors protects brain against injury in a rat model of focal cerebral ischemia." in: Journal of neuroinflammation, Vol. 3, pp. 12, (2006) (PubMed).

Target Details

Target: Glucose

Background: Quantitative determination of glucose by chemical colorimetric (630nm) method.
Procedure: 10 min.

Glucose (C6H12O6) is a ubiquitous fuel molecule in biology. It is oxidized through a series of enzyme-catalyzed reactions to form carbon dioxide and water, yielding the universal energy molecule ATP. Due to its importance in metabolism, glucose level is a key diagnostic parameter for many metabolic disorders. Increased glucose levels have been associated with diabetes mellitus, hyperactivity of thyroid, pituitary and adrenal glands. Decreased levels are found in insulin secreting tumors, myxedema, hypopituitarism and hypoadrenalism. Simple, direct and automation-ready procedures for measuring glucose concentrations find wide applications in research and drug discovery. This glucose assay kit is designed to measure glucose directly in serum or plasma without any pretreatment. The improved o-toluidine method utilizes a specific color reaction with glucose. The absorbance at 630nm is directly proportional to glucose concentration in the sample.