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By Michael Williams (auth.), Michael Williams (eds.)

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As would be expected for an antagonist radioligand, agonists were about five times less potent in displacing PD115,199, whereas antagonists were three times more potent than comparable data obtained using PH]NECA in the presence of CPA. The stability of this xanthine radioligand, however, is poor. 2. , 1982), a triazolquinazoline whose predominant activity was at the benzodiazepine receptor. Subsequent SAR work on this novel heterocycle led to the identification of CGS 15943 (Fig. , 1987a). Labeling of this adenosine receptor antagonist has been confounded by ligand instability.

251,279-287 Barrington, W. , Jacobson, K. , and Stiles, G. L. (1989a) Demonstration of distinct agonist and antagonist conformations of the Al adenosine receptor. I. Bioi. Chem. 264, 13157-13164. Barrington, W. , Jacobson, K. , Hutchison, A. , and Stiles, G. L. (l989b) Identification of the A,. adenosine receptor binding subunit by photoaffinity crosslinking. Proc. Nail. Acad. Sci. USA 86, 6572-6576. Beck, D. , Vinters, H. , Moore, S. , Hart, M. , Henn, F. , and Cancilla, P. A. (1984) Demonstration of adenosine receptors on mouse cerebral smooth muscle membmnes.

20 nM), fIltration is an effective and reliable means by which to separate bound radioactivity from free. As the affInity decreases (an increase in the Kd value), fIltration tends to remove bound as well as free radioactivity, leading to an underestimate of both parameters and providing extremely variable data. This problem can be circumvented by the use of centrifugation to pellet the receptor-ligand complex. This generally allows a more reliable estimate of radioactivity bound with low affInity, but is also more time consuming and results in higher background radioactivity.

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