Saturation Binding Lab Report

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The following report investigates the analytical method that are used in the determination of the binding properties of 3H–MPPF in a rat hippocampus. The method used in the following report is saturation binding. The report also examines the data interpretation of the results that are obtained and compares the old technique of Scathard plots to the new analysis that may be conducted on graph pad prism. The saturation binding experiment is based on the law of mass action and therefore it is important to know the ideas upon which this model works. The following are the principle foundation in the law of mass action.
I. Binding occurs because of the ligand having the correct orientation to the bind to the receptor in the presence of sufficient
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After binding has completed the receptor and ligand complex will remain bound for a period of time, which is influenced by the affinity of the ligand for the receptor.
III. After association of the ligand receptor complex has completed the ligand and receptor will remain unchanged.
IV. Analysis is performed when equilibrium is obtained. In this circumstance, it refers to the association rate and dissociation rate of complex formation being equal. The fractional occupancy at equilibrium is predicted by the law of mass action
Firstly, a protein standard curve will be created to determine the concentration of the unknown sample. When this is completed, the saturation binding graph will be created using graph pad prism and determine the Bmax and Kd values. A Scathard plot will then be created and the Bmax obtained from the x-axis intercept and the Kd value obtained from the slope of the line the. The results will be discussed and the difference between the graphs values obtained will be reviewed.
Total binding of the radioactive drug to the membrane will be obtained. This consists of both non-specific and specific binding. Further details on what non-specific binding is and how it is overcome in analysis can be found under the exercise
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