Journal article

BIOCHEMICAL-CHARACTERIZATION OF AN AUTORADIOGRAPHIC METHOD FOR STUDYING EXCITATORY AMINO-ACID RECEPTORS USING L-[H-3]GLUTAMATE

M CINCOTTA, RJ SUMMERS, PM BEART

ANALYTICAL BIOCHEMISTRY | ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS | Published : 1989

Abstract

A method was developed for radiolabeling excitatory amino acid receptors of rat brain with L-[3H]glutamate. Effective labeling of glutamate receptors in slide-mounted 10-microns sections was obtained using a low incubation volume (0.15 ml) and rapid washing: a procedure where high ligand concentrations were achieved with minimal waste. Saturation experiments using [3H]glutamate revealed a single binding site of micromolar affinity. The Bmax was trebled in the presence of Ca2+ (2.5 mM) and Cl- (20 mM) with no change in the Kd. Binding was rapid, saturable, stereospecific, and sensitive to glutamate receptor agonists. The proportions of [3H]glutamate binding sensitive to N-methyl-D-aspartate (..

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