I would like to measure dexamethasone binding affinity in PBMC and due to bureaucracy I would like to avoid using labeled radioactive DEX . Does anybody know of any other possible protocol?
we don't have a binding affinity test, but we do have an IC50 test that gives a good readout of GR functioning.
We stimulate PBMC's with LPS in the presence of a range of Dex concentrations, and measure IL6 secretion. From the IL6 curve, we calculate the IC50 of Dex, ie the dex concentration that reduces by 50% the IL6 secretion.
Don't know if you can adapt this, but it should give you an idea of how much *functional" GR you have.