PCL475Y1 Lecture Notes - Lecture 9: Lipid Bilayer, Area Postrema, Dissociation Constant
28 Aug 2019
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Binding assay: homogenize break down tissue, breaking down cells, spin it down to get rid of cytoplasm and nuclear material. Drug + receptor drug/receptor complex (low kd, saturable) Drug + lipid drug/lipid complex (no real binding, no kd, non-saturable) Actual filter assay (two tubes: use radioactivity to destroy the receptor bound to drug (expensive and difficult, use test tube: One has hot drug receptors and overabundance of cold drug. Saturation is expensive (use of a lot of hot ligand to establish basic parameters of binding) Competition is cheaper (what other compounds bind to this site?) Proves what other ligands bind to this receptors other than your hot ligand. Saturation experiments give: kd concentration of hot drug that binds half the receptors, bmax number of receptors that are there to be bound to, dose response curve or concentration binding curve. The whole animal response reflects response of drug binding to receptors.
