Elimination of glucose contamination from adipocyte glycogen extracts

• Autor(a) principal: Nunes, Patricia M.
• Data de Publicação: 2008
• Outros Autores: Carvalho, Eugénia, Jones, John G.
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10316/5289; https://doi.org/10.1016/j.carres.2008.04.004
• topic: Adipocyte; Glycogen extraction; Glucose oxidase; Amyloglucosidase

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.