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Download e-book for kindle: Brain Receptor Methodologies. Part A: General Methods and by Paul J. Marangos

By Paul J. Marangos

ISBN-10: 012470350X

ISBN-13: 9780124703506

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Extra resources for Brain Receptor Methodologies. Part A: General Methods and Concepts. Amines and Acetylcholine

Example text

J. Neurochem. 35, 5 - 1 5 . Thakur, A. , Jaffe, M. , and Rodbard, D. (1980). Anal. Biochem. 107, 2 7 9 - 2 9 5 . -F. (1975). " Academic Press, New York. Chapter 4 THE DISTRIBUTION OF PERIPHERAL REGULATORY PEPTIDES: A DUAL IMMUNOCHEMICAL (IMMUNOCYTOCHEMISTR Y AND RADIOIMMUNOASSAY) APPROACH JULIA M. POLAK Department of Histochemistry Hammersmith Hospital Royal Postgraduate Medical School London, England STEPHEN R. BLOOM Department of Medicine Hammersmith Hospital Royal Postgraduate Medical London, England I.

Tritium and I are the two radionuclides most commonly used to achieve this specific activity. Accordingly, this chapter will be limited to methods utilized to introduce tritium 1 2 5 and I labels. Generally, the ligand binding study is carried out so that less than 10% of the total radioligand is bound at equilibrium (Bennett, 1978). As a result, the radiochemical purity of the ligand is of utmost importance; a minimum purity of 98% is usually required. In order to maintain this level of purity for an acceptable period of time, the stability of the radioligand is of concern.

Thus, determination of receptor homogeneity must go beyond statements about the linearity of the Scatchard plot. Competition studies (in which varying doses of unlabeled ligand compete with labeled species for a finite number of receptor binding sites) may also reveal receptor heterogeneity. Reduced slope of the competition curve or inability to completely displace labeled ligand suggest the presence of multiple classes of receptors. Interestingly, saturation studies and competition studies do not really offer independent evidence of receptor heterogeneity, since both studies measure binding of varying hormone concentrations to the same receptor preparation.

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Brain Receptor Methodologies. Part A: General Methods and Concepts. Amines and Acetylcholine by Paul J. Marangos


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