Methodology for benzodiazepine receptor binding assays at physiological temperature. Rapid change in equilibrium with falling temperature
Benzodiazepine receptors of rat cerebellum were assayed with (/sup 3/H)-labeled flunitrazepam at 37/sup 0/C, and assays were terminated by filtration in a cold room according to one of three protocols: keeping each sample at 37 degrees C until ready for filtration, taking the batch of samples (30) into the cold room and filtering sequentially in the order 1-30, and taking the batch of 30 samples into the cold room and filtering sequentially in the order 30-1. the results for each protocol were substantially different from each other, indicating that rapid disruption of equilibrium occurred as the samples cooled in the cold room while waiting to be filtered. Positive or negative cooperativity of binding was apparent, and misleading effects of gamma-aminobutyric acid on the affinity of diazepam were observed, unless each sample was kept at 37/sup 0/C until just prior to filtration.
- Research Organization:
- Materials Research Labs., Ascot Vale, Victoria, Australia
- OSTI ID:
- 6842114
- Journal Information:
- J. Pharmacol. Methods; (United States), Vol. 4
- Country of Publication:
- United States
- Language:
- English
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TRANQUILIZERS
RADIORECEPTOR ASSAY
CEREBELLUM
FILTRATION
RATS
RECEPTORS
TEMPERATURE DEPENDENCE
TRITIUM COMPOUNDS
ANIMALS
BODY
BRAIN
CENTRAL NERVOUS SYSTEM
CENTRAL NERVOUS SYSTEM AGENTS
DRUGS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
NERVOUS SYSTEM
ORGANIC COMPOUNDS
ORGANS
PROTEINS
PSYCHOTROPIC DRUGS
RODENTS
SEPARATION PROCESSES
TRACER TECHNIQUES
VERTEBRATES
551001* - Physiological Systems- Tracer Techniques