I'm using a glucose colorimetric detection kit from Thermo (EIAGLUC), but I always get a lot of intra and inter-assay variability, sometimes even between replicates. I'm measuring glucose in culture media after 10 minutes of insulin stimulated uptake. The kit protocol misses a lot of information and does not include troubleshooting, so I don't know if that kit is not good, if it is not enough time for the tissue to capture glucose or if the problem is human error.
If you could recommend a kit to detect glucose that uses fluorescence I would really appreciate it.
Thank you!
If you would like to evaluate the uptake of glucose into cells as in the following paper, you should better see the amount of 2-deoxyglucose in the cells.
Article Impact of Divergent Effects of Astaxanthin on Insulin Signal...
Because.
1. Because you will measure slight changes from extremely high concentrations, the amount of glucose in the culture medium you measure is expected to vary widely, confounded by measurement error.
2. the amount of glucose in the cells is metabolized in a very short time.
For these reasons, it is appropriate to evaluate cellular glucose uptake by quantifying glucose analogues such as labeled 2-deoxyglucose.
Please note that the evaluation is based on cell lysates, not culture medium. The kit is shown below. Please refer to them.
https://www.promega.jp/products/energy-metabolism/metabolite-detection-assays/glucose-uptake_glo-assay?gclid=CjwKCAjwquWVBhBrEiwAt1KmwsARha9kqOQkpWnNPqkm1Orfh2CyQqIdW-uZIwfxGaYliJoImW9HHBoCKK4QAvD_BwE&catNum=J1341
Good luck！
Yasuhiro Nishida The thing is that the experiment is finished and I no longer have the tissue, just the culture media I stored, so I cannot incubate them with 2-DG. I need something that doesn't involve culturing again, just to measure the amount of glucose in the medium.
Thank you so much for your recommendation I will consider it for the next experiment!
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