3T3-L1 cells are insulin sensitive but when they are counted and seeded for insulin stimulated glucose uptake assay, it results with 123% of glucose being up-taken by the positive control cells treated with 100 nM insulin. Can the % of glucose uptake be more than 100%?
Hey,
How do you do your calculations? Do you youse 3H labelled 2DOG or a different compound?
Depends on your margin of error in quantitation and how you calculate (normalization)!
Post details and you will most likely get better quality help.
Hi Karin, I'm not using radio-labelled 2DOG. I m following enzymatic 2DG uptake assay using glucose oxidase.
@abishek: thank u very much sir and my standard concentration is 1.64mg/ml which is assumed as 100 and my calculations give me 98% uptake approximately for positive control.
Yes, its conventional method. And thank you for your suggestions. I will check it out
ah, I see, I have never done that, sorry. In my method I trace medium of known glucose concentration with radioactive 2DOG, measure activity per volume in this solution, and from that can calculate back which volume (and thus amount of glucose) the measured radioactivity per count corresponds to. Good luck with your Assay!
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