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The Hill equation reflects the occupancy of macromolecules: the fraction that is saturated or bound by the ligand. [1] [2] [nb 1] This equation is formally equivalent to the Langmuir isotherm. [3] Conversely, the Hill equation proper reflects the cellular or tissue response to the ligand: the physiological output of the system, such as muscle ...
A ligand binding assay (LBA) is an assay, or an analytic procedure, which relies on the binding of ligand molecules to receptors, antibodies or other macromolecules. [1] A detection method is used to determine the presence and amount of the ligand-receptor complexes formed, and this is usually determined electrochemically or through a fluorescence detection method. [2]
The first description of cooperative binding to a multi-site protein was developed by A.V. Hill. [4] Drawing on observations of oxygen binding to hemoglobin and the idea that cooperativity arose from the aggregation of hemoglobin molecules, each one binding one oxygen molecule, Hill suggested a phenomenological equation that has since been named after him:
Rotating cell‑based ligand binding assay using radioactivity or fluorescence, is a recent method that measures molecular interactions in living cells in real-time. This method allows the characterization of the binding mechanism, as well as K d, k on and k off. This principle is being applied in several studies, mainly with protein ligands ...
Hill equation [ edit ] A simple and widely used model for molecular interactions is the Hill equation , which provides a way to quantify cooperative binding by describing the fraction of saturated ligand binding sites as a function of the ligand concentration.
For competition binding assays and functional antagonist assays IC 50 is the most common summary measure of the dose-response curve. For agonist/stimulator assays the most common summary measure is the EC 50. [6] The EC 50 is also related to IC 50 which is a measure of a compound's inhibition (50% inhibition).
Several reliable methods for quantifying protein have been developed to simplify the process. These methods include Warburg–Christian method, Lowry assay, and Bradford assay (all of which rely on absorbance properties of macromolecules). Bradford assay method uses a dye to bind to protein. Most commonly, Coomassie brilliant blue G-250 dye is ...
The Hill equation can be used to describe dose–response relationships, for example ion channel-open-probability vs. ligand concentration. [9] Dose is usually in milligrams, micrograms, or grams per kilogram of body-weight for oral exposures or milligrams per cubic meter of ambient air for inhalation exposures. Other dose units include moles ...
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