Cells uptake glucose based on their metabolic needs. If they are entering a more active state, they will have a higher energy requirement and therefore more glucose will be taken up from the extracellular environment. Measurement of glucose uptake is crucial in a range of research areas including cancer, diabetes and immunology.
Traditional methods for measuring glucose uptake in cultured cells used glucose analogues. These analogues, such as 2deoxyglucose (2-DG), are taken up by the cell in the same way as glucose (through GLUT transporters on the cell membrane) but are not metabolised intracellularly in the same way as glucose. This means that they accumulate in the cell and the window of detection for the researcher is very narrow. The best way would be to be able to directly measure the natural glucose which the cell takes in.
This is where Assay Genie’s new Direct Glucose Uptake Assay kit comes in. This kit measures the physiological Glucose Uptake and has a less than 15% variability versus 60% with 2-DG methods. A specific inhibitor is used to inhibit hexokinase activity, the first enzyme which metabolises glucose in the cell. Enzymatic reactions oxidise the glucose to intermediates which react with a fluorescent probe from the kit, resulting in a directly proportional fluorescent signal to the amount of glucose taken up. The assay is also super-sensitive, capable of measuring as little as 5 pmol, and really easy to use.