Glycerol Assay Kit

Produktnummer ABIN1000312

Produktdetails

Target: Glycerol

Applikation: Biochemical Assay (BCA)

Proben: Serum, Plasma

Spezifität: 2 μM

Produktmerkmale: Sensitive and accurate. Use as little as 10 µL samples. Linear detection range in 96-well plate: 10 to 1000 µM (92 µg/dL to 9.2 mg/dL) glycerol for colorimetric assays and 2 to 50 µM for fluorimetric assays.
Simple and convenient. The procedure involves addition of a single working reagent and incubation for 20 min at room temperature, compatible for HTS assays.
Improved reagent stability. The optimized formulation has greatly enhanced the reagent and signal stability.

Bestandteile: Assay Buffer: 24 mL. Enzyme Mix: 500 µL. ATP: 250 µL. Dye Reagent: 220 µL. Standard: 100 µL 100 mM Glycerol.

Benötigtes Material: Pipeting devices, centrifuge tubes, Clear flat-bottom 96-well plates, black 96-well plates (e.g. Corning Costar) and plate reader.

Anwendungsinformationen

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

Protokoll: Note: SH-group containing reagents (e.g. mercaptoethanol, DTT) may interfere with this assay and should be avoided in sample preparation.
1. Equilibrate all components to room temperature. Keep thawed Enzyme Mix in a refrigerator or on ice. Dilute standard in distilled water (diluted standards can be used for future assays when stored refrigerated). Transfer 10 µL standards and 10 µL samples into separate wells of a clear 96-well plate.
2. For each reaction well, mix 100 µL Assay Buffer, 2 µL Enzyme Mix, 1 µL ATP and 1 µL Dye Reagent in a clean tube. This Working Reagent should be used on the same day of preparation. Transfer 100 µL Working Reagent into each reaction well. Tap plate to mix.
3. Incubate 20 min at room temperature. Read optical density at 570nm (550-585nm). Note: if the Sample OD is higher than the Standard OD at 1.0 mM, dilute sample in water and repeat the assay. Multiply result by the dilution factor.

Ergebnisberechnung:

Subtract blank OD (water, #4) from the standard OD values and plot the OD against standard concentrations.
Conversions: 1mM glycerol equals 9.2 mg/dL, 92 ppm. FLUORIMETRIC

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C

Referenzen

Calamita, Gena, Ferri, Rosito, Rojek, Nielsen, Marinelli, Frühbeck, Svelto: "Biophysical assessment of aquaporin-9 as principal facilitative pathway in mouse liver import of glucogenetic glycerol." in: Biology of the cell / under the auspices of the European Cell Biology Organization, Vol. 104, Issue 6, pp. 342-51, (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).

Wächtler, Wilson, Haedicke, Dalle, Hube: "From attachment to damage: defined genes of Candida albicans mediate adhesion, invasion and damage during interaction with oral epithelial cells." in: PLoS ONE, Vol. 6, Issue 2, pp. e17046, (2011) (PubMed).

Pifferi, Tremblay, Croteau, Fortier, Tremblay-Mercier, Lecomte, Cunnane: "Mild experimental ketosis increases brain uptake of 11C-acetoacetate and 18F-fluorodeoxyglucose: a dual-tracer PET imaging study in rats." in: Nutritional neuroscience, Vol. 14, Issue 2, pp. 51-8, (2011) (PubMed).

Bahar, ODoherty, Sweeney: "A potential role of IL-6 in the chito-oligosaccharide-mediated inhibition of adipogenesis." in: The British journal of nutrition, Vol. 106, Issue 8, pp. 1142-53, (2011) (PubMed).

Lung, Shulga, Ivanova, Myers, Milne, Brown, Topham, Epand: "Diacylglycerol kinase epsilon is selective for both acyl chains of phosphatidic acid or diacylglycerol." in: The Journal of biological chemistry, Vol. 284, Issue 45, pp. 31062-73, (2009) (PubMed).

Antigendetails

Target: Glycerol

Substanzklasse: Alcohol

Hintergrund: Quantitative determination of glycerol by colorimetric (570nm) or fluorimetric (530nm/590nm) methods.
Procedure: 20 min.

Glycerol [Glycerin or Glycerine, C3H5(OH)3] is widely used in foods, beverages and pharmaceutical formulations. It is also a main by-product of biodiesel production. Simple, direct and automation-ready procedures for measuring glycerol concentrations find wide applications. This glycerol assay uses a single Working Reagent that combines glycerol kinase, glycerol phosphate oxidase and color reactions 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 glycerol concentration in the sample.