A high-performance liquid chromatography assay of brain adenylate cyclase using [3H]ATP as substrate
- 1 February 1986
- journal article
- research article
- Published by Springer Nature in Neurochemical Research
- Vol. 11 (2) , 161-171
- https://doi.org/10.1007/bf00967965
Abstract
A sensitive, reproducible assay for adenylate cyclase is described which separates labeled cyclic AMP from ATP and other nucleotides by high-performance liquid chromatography (HPLC) on reverse-phase columns. The technique utilizes [3H]ATP as substrate, and the principal compound contaminating the [3H]cyclic AMP peak, adenosine, is removed by incubation of assay tubes with small amounts of adenosine deaminase. The HPLC elution utilizes high resolution (3 μm) short (10 cm) C-18 columns for increased resolution and decreased flow rates. Since cyclic AMP elutes at 4 min following injection, this procedure can easily process large numbers of samples per day when combined with automated techniques of sample injection and collection.This publication has 17 references indexed in Scilit:
- An Improved, Automated Adenylate Cyclase Assay Utilizing Preparative HPLC: Effects of Phosphodiesterase InhibitorsJournal of Neurochemistry, 1984
- A modified assay of adenylate cyclase using 3H and 14CAnalytical Biochemistry, 1983
- Activation of adenylate cyclase by the diterpene forskolin does not require the guanine nucleotide regulatory protein.Journal of Biological Chemistry, 1981
- The role of hormone receptors and GTP-regulatory proteins in membrane transductionNature, 1980
- Agonist-promoted coupling of the beta-adrenergic receptor with the guanine nucleotide regulatory protein of the adenylate cyclase system.Proceedings of the National Academy of Sciences, 1980
- Improved two-step method for the assay of adenylate and guanylate cyclaseAnalytical Biochemistry, 1978
- A highly sensitive adenylate cyclase assayAnalytical Biochemistry, 1974
- An improved protein binding assay for cyclic AMPAnalytical Biochemistry, 1974
- A simple and sensitive saturation assay method for the measurement of adenosine 3′:5′-cyclic monophosphateBiochemical Journal, 1971
- The role of cyclic-3',5'-AMP in responses to catecholamines and other hormones.1966