Elimination of glucose contamination from adipocyte glycogen extracts

• Author(s): Nunes, Patricia M. ; Carvalho, Eugénia ; Jones, John G.
• Date: 2008
• Persistent ID: http://hdl.handle.net/10316/5289
• Origin: Estudo Geral - Universidade de Coimbra
• Subject(s): Adipocyte; Glycogen extraction; Glucose oxidase; Amyloglucosidase
• Document Type: Article
• Language: English

Description

Glycogen was quantified in rat adipocytes by isolation using conventional KOH digestion and ethanol precipitation, followed by hydrolysis and spectrophotometric assay of the glucose product. A concentration of 0.193 ± 0.020 [mu]mol glucosyl units/106 cells was recorded. When this procedure was modified by including a 4 h incubation with glucose oxidase prior to glycogen hydrolysis, the glycogen concentration was found to be 0.055 ± 0.008 [mu]mol glucosyl units/106 cells. Therefore in adipocytes, conventional glycogen assays give substantial overestimates due to incomplete removal of glucose during glycogen isolation. Contaminant glucose can be scavenged in a simple manner by incubation with glucose oxidase prior to glycogen hydrolysis. http://www.sciencedirect.com/science/article/B6TFF-4S7906X-4/1/a08b7a48b48b25f3f821340d39b80c8f