Glucose uptake assay kit

The glucose uptake by LX2 cells was determined using a Glucose Uptake Assay Kit (Abnova, Taiwan, China) according to the manufacturer’s instructions. In this assay, the glucose analog 2-DG is metabolized to 2-DG-6-phosphate, which is proportional to glucose uptake by cells. The accumulated 2-DG-6-phosphate is enzymatically coupled to generate NADPH, which is specifically monitored by a NADPH sensor. The signal can be read by an absorbance microplate reader by reading the OD ratio at wavelength 570 to 610 nm.

Levels of adenosine triphosphate (ATP) (A095) and lactic acid (A019-02), as well as glucose uptake (cat. KA4086) in B-CPAP cells, were evaluated using corresponding commercial kits. The ATP and lactic acid assay kits were supplied by the Nanjing Jiancheng Bioengineering Institute. The glucose uptake assay kit was supplied by Abnova.

Glucose Uptake Assay Kit (#KA4086, Abnova, Taiwan) was used for glucose uptake measuring. First, 5 × 10⁴ cells/well/100 μL/were seeded in a 96-well plate in treatment medium which containing 0.3% DMSO (0 μM P7C3), 30 μM P7C3, or 50 μM P7C3 for 18 h. The medium was then replaced with a new growth medium, and the cells were cultured for another 6 hours before experiments. Then, the cells were treated as desired, and 10 μL/well 2-Deoxy-d-glucose was added, and the cells were incubated at 37°C for 30 min. To wash and lyse cells, 50 μL/well 2-Deoxy-d-glucose uptake assay working solution was added, and the cells were incubated at RT for 90 min. The optical density (OD) ratio at 570/610 nm was measured using a spectrophotometer (1510, Thermo Fisher, USA).