Glucose Assay Kit

Catalog No. ABIN1000294

Product Details

Target: Glucose

Application: Biochemical Assay (BCA)

Sample Type: Beverages, Cell Culture Supernatant, Food, Milk, Plasma, Saliva, Serum, Urine

Specificity: 5 μM

Characteristics: Sensitive and accurate. Use as little as 20 µL samples. Linear detection range in 96-well plate: 5 to 300 µM (90 µg/dL to 5.4 mg/dL) glucose for colorimetric assays and 1 to 30 µM for fluorimetric assays.
Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 30 min at room temperature.

Components: Assay Buffer: 10 mL. Enzyme Mix: 120 µL. Dye Reagent: 120 µL. Standard: 1 mL 300 mg/dL Glucose.

Material not included: Pipetting devices, centrifuge tubes, clear flat-bottom 96-well plates, black 96-well plates and plate reader.

Application Details

Application Notes: Direct Assays: glucose in serum, plasma, urine, saliva, milk, culture medium and other biological samples.
Drug Discovery/Pharmacology: effects of drugs on glucose metabolism.
Food and Beverages: glucose in food, beverages etc.

Protocol: 1. Equilibrate all components to room temperature. During experiment, keep thawed Enzyme in a refrigerator or on ice.
2. Standards and samples: for 300 µMstandard, mix 15 µL 300 mg/dL standard with 818 µL dH2O. Dilute standard in dH2O. No 300 µMSTD + H2O Vol (µL) Glucose (µM) 1 200 µL + 0 µL 200 300 2 120 µL + 80 µL 200 180 3 60 µL + 140 µL 200 90 4 0 µL + 200 µL 200 0 Transfer 20 µL standards and samples into separate wells.
3. Working Reagent. For each reaction well, mix 85 µL Assay Buffer, 1 µL Enzyme Mix (vortex briefly before pipetting), and 1 µL Dye Reagent in a clean tube. Transfer 80 µL Working Reagent into each reaction well. Tap plate to mix.
4. Incubate 30 min at room temperature. Read optical density at 570nm (550-585nm).

Sample Preparation:

Saliva samples should be centrifuged for 5 min at 14,000 rpm prior to assay. Milk samples should be cleared by mixing 100 µL 6N HCl and 600 µL milk. Centrifuge 5 min at 14,000 rpm and transfer supernatant into a clean tube. Add 170 µL 6N NaOH per mL supernatant. Mix well and centrifuge again at 14,000 rpm. The supernatant can be assayed. The dilution factor in this procedure is n = 1.36. Samples can be analyzed immediately after collection, or stored in aliquots at -20 °C. Avoid repeated freeze-thaw cycles. If particulates are present, centrifuge sample and use clear supernatant for assay.

Calculation of Results:

Subtract blank OD (water, #4) from the standard OD values and plot the OD against standard concentrations.
Conversions: 1 mg/dL glucose equals 55.5 µM, 0.001% or 10 ppm. FLUORIMETRIC

Restrictions: For Research Use only

Handling

Storage: -20 °C

References

He, Meng, Germain-Lee, Radovick, Wondisford: "Potential biomarker of metformin action." in: The Journal of endocrinology, (2014) (PubMed).

Klippel, Jakubikova, Delmore, Ooi, McMillin, Kastritis, Laubach, Richardson, Anderson, Mitsiades: "Methyljasmonate displays in vitro and in vivo activity against multiple myeloma cells." in: British journal of haematology, Vol. 159, Issue 3, pp. 340-51, (2012) (PubMed).

Drew, Carey, Natoli, Formosa, Vizi, Reddy-Luthmoodoo, Weir, Barlow, van Hall, Meikle, Duffy, Kingwell: "Reconstituted high-density lipoprotein infusion modulates fatty acid metabolism in patients with type 2 diabetes mellitus." in: Journal of lipid research, Vol. 52, Issue 3, pp. 572-81, (2011) (PubMed).

Dhar, Desai, Wu: "Alagebrium attenuates acute methylglyoxal-induced glucose intolerance in Sprague-Dawley rats." in: British journal of pharmacology, Vol. 159, Issue 1, pp. 166-75, (2010) (PubMed).

Cha, Wolfgang, Tokutake, Chohnan, Lane: "Differential effects of central fructose and glucose on hypothalamic malonyl-CoA and food intake." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, Issue 44, pp. 16871-5, (2008) (PubMed).

Wolfgang, Cha, Sidhaye, Chohnan, Cline, Shulman, Lane: "Regulation of hypothalamic malonyl-CoA by central glucose and leptin." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, Issue 49, pp. 19285-90, (2007) (PubMed).

Wolfgang, Kurama, Dai, Suwa, Asaumi, Matsumoto, Cha, Shimokawa, Lane: "The brain-specific carnitine palmitoyltransferase-1c regulates energy homeostasis." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, Issue 19, pp. 7282-7, (2006) (PubMed).

Target Details

Target: Glucose

Background: Quantitative determination of glucose by colorimetric (570nm) or fluorometric (530nm/590nm) methods.
Procedure: 40 min.

Glucose (C6H12O6) 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 high-throughput assays for measuring glucose concentrations find wide applications in research and drug discovery. This glucose assay kit uses a single Working Reagent that combines the glucose oxidase reaction and color reaction in one step. The color intensity of the reaction product at 570nm or fluorescence intensity at gamma em/ex = 585/530nm is directly proportional to glucose concentration in the sample.