By Paul J. Marangos
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Additional resources for Brain Receptor Methodologies. Part A: General Methods and Concepts. Amines and Acetylcholine
High values of (BIF)0 are not uncommon when using high-affinity, partially purified receptors, hence further correction of the results may be necessary. The exact relationship of the Kx to the E D 5 0 is given in Rodbard and Lewald (1970). A superior approach, which does not make use of any of the above approximations, is the model-based curve-fitting method. Rather than estimating the E D 5 0 3. Heterogeneous Receptors and Binding Curve Analysis in Neurobiology 43 values for each curve independently and then converting these to Kx estimates, it is possible, with computerized curve fitting, to estimate the Kx directly from the data.
Therefore, any tendency toward a linear trend ought to signal the presence of saturable binding. Unfortunately, in all but the most clearcut cases, the distortions of statistical errors resulting from the data transformation and the plotting technique itself make this approach unworkable. The Scatchard plot method may be subject to serious misinterpretation if, in addition to a linear trend, there is a suggestion of curvature in the data. Finally, the definition of saturability depends strongly upon how the nonspecific binding is defined and measured.
In order to prevent such difficulties, the investigator should become familiar with the statistical principles guiding t-»e analysis. Computerized analyse (in fact, any precise analytic method) begins with the postulation of a matherr atical or biochemical model or framework into which the 3. Heterogeneous Receptors and Binding Curve Analysis in Neurobiology 37 experimental data are fitted. The model may take the form of a set of equations or may be defined by the computer program actually used in calculation.