I'm trying to measure the extracellular glucose of my cells to get a feel for their rate of glucose uptake (from a fixed concentration of glucose), however I'm not really an experimentalist and am struggling to find a protocol for this. Our lab is equipped with spectrophotometers and all major enzymes and co-factors of the glycolytic pathway.
Hello Stefan, you can use glucose oxidase-peroxidase (GOD/POD) method for direct estimation of glucose from media or you can use fluorescent labeled glucose (2-NBDG) for indirect estimation of glucose uptake from media. both methods are quite sensitive and accurate for glucose uptake measurement. just measure the glucose in sample before incubation and after incubation.
It is much better to measure the glucose in the cell. Not media. Remember the medium is equivalent to a ocean and the cells are like a thin scum of algae. By looking at medium, you are lowering sensitivity of assay alot.
You can measure the uptake of 2 deoxy glucose by spectrophotometry using available kits.
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