6533b839fe1ef96bd12a65bc
RESEARCH PRODUCT
A Microassay for Measuring Glycogen in 96-Well-Cultured Cells
J.v. CastellMaria-josé Gómez-lechónXavier Ponsodasubject
MaleTime FactorsBiophysicsSensitivity and SpecificityBiochemistryRats Sprague-DawleyHydrolysischemistry.chemical_compoundCarbohydrate ConformationAnimalsGlucose oxidaseMolecular BiologyCells CulturedSample handlingchemistry.chemical_classificationChromatographybiologyGlycogenHydrolysisMicrochemistryfungiColor intensityRapid processingReproducibility of Resultsfood and beveragesDNACell BiologyLiver GlycogenRatsGlucoseEnzymeLiverBiochemistrychemistrybiology.proteinColorimetryGlucan 14-alpha-GlucosidaseIntracellulardescription
Abstract This study describes a rapid, sensitive, and automated spectrophotometric enzymatic microassay that measures the intracellular glycogen of primary cultured hepatocytes and other cultured cells in 96-well plates and can be adapted for other samples that are transferred to these plates. The procedure involves in situ disruption of cells, followed by hydrolysis of glycogen into glucosyl units by fungal glucoamylase (exo-1,4-α- D -glucosidase, EC 3.2.1.3), and glucose determination with the glucose oxidase colorimetric method. The color intensity can be measured in conventional ELISA readers, and the data can be fed to an on-line computer for rapid processing. The advantages of this method are its simplicity and automation, the reduction in sample handling, and the small number of cells required compared to other conventional methods.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1996-05-01 | Analytical Biochemistry |