I am performing a glucose uptake assay in differentiated C2C12 cells using [3H]2-deoxyglucose. The protocol that I am using was adapted from a glucose uptake protocol optimized for 3T3-L1 cells. In this protocol they glucose-starve the 3T3-L1 cells in KRH buffer with 2.5 mM sodium pyruvate and 0.5% BSA. From my review of the literature, sodium pyruvate is not normally added for C2C12 cells.
Therefore, I wanted to know whether I should add sodium pyruvate (2.5 mM) to my KRH buffer when I starve my C2C12 cells of glucose for 2h before the glucose uptake assay?
Hi Goergia,
Since you will be performing serum starvation for 2 hrs only, you can skip addition of sodium pyruvate to the KRH buffer. I have performed glucose uptake assay on 3T3L1 with sodium pyruvate.
All the best.
Regards,
Prasad
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