hello ，I want to use exogenous ligands to activate receptors on the surface of the U87 cell line. How can I remove endogenous ligands before the experiment to ensure a low baseline receptor activation level? I tried treating the cells with a glycine buffer at pH 2.7 (30 seconds * 3 times), but after treatment, my cells died. How can I optimize my experimental conditions? thanks for your kindly help.
Adam B Shapiro
As long as the affinity of the ligands is not too high, simple dilution should allow them to dissociate. Remove the medium containing the ligand and replace it with a large volume of fresh medium. Allow some time (half an hour?) for dissociation. Repeat this process a few times.
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