I am trying to measure insulin secretion from HIT-T15 cell lines using ELISA.
To culture the cell line, I am using Hank's F12K as a basal media, but it contains 10mM of glucose, and I cannot find the media with low glucose.
From the protocol that I am referring to, they incubated beta cells in low glucose media for 5 hours for an adaptation process, and change with flesh low glucose media for 2 hours before running ELISA with the media.
How would you do? Do you know how to prepare Hank's F12k from powders?
or Can I use any buffer solution to measure insulin?
DMEM and RPMI are available without any added glucose. To test the effect of various concentrations of glucose, prepare a 2M stock solution, filter sterilize it and add the appropriate volume.
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