Glycogen Content Measurement in Myotubes

Glycogen content was measured as described in by Cagin, U. et al. [75 (link)]. The glycogen was indirectly measured whilst the glucose was released after total digestion with amyloglucosidase from Aspergillus niger (Merck Life Science, Darmstadt, Germany). Sol8 myoblasts were differentiated into myotubes, as previously described. Lysates from cells and animal tissues were incubated for 5 min at 95 °C and then cooled at 4 °C; 25 μL of amyloglucosidase diluted at 1:5 in 0.1M potassium acetate (pH 5.5) was added to each sample. A control reaction without amyloglucosidase was prepared for each sample. Both sample and control reactions were incubated at 37 °C for 90 min. The reaction was stopped by incubating samples for 5 min at 95 °C. The released glucose was determined using a glucose assay kit (Merck Life Science) by measuring absorbance with Infinite 200 PRO NanoQuant (Tecan, Männedorf, Switzerland) at 540 nm.