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Comparison of centrifugation and filtration assays of ligand binding: Do multiple GABA receptive sites exist?
Author(s) -
Patel Shutish C.,
Peck Ernest J.
Publication year - 1982
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490080405
Subject(s) - muscimol , filtration (mathematics) , chemistry , membrane , centrifugation , chromatography , ligand (biochemistry) , gabaa receptor , ligand binding assay , receptor , binding site , biophysics , biochemistry , biology , mathematics , statistics
Filtration and microcentrifugation procedures for the separation of bound and free ligand are compared for the assay of GABA receptors. The 3 H‐GABA and 3 H‐muscimol are employed as ligands in saturation analyses of SPM and JC preparations. A direct comparison of the two methods, applied to two membrane preparations and two ligands, reveals that nonspecific binding is consistently higher with the microcentrifugation procedure. Analysis of the binding data yields essentially the same constants in either case; however, the filtration assay provides a better estimate in all cases.