High-performance liquid chromatography assay of brain adenylate cyclase using (/sup 3/H)ATP as substrate

Journal Article · Fri Jan 31 23:00:00 EST 1986 · Neurochem. Res.; (United States)

DOI:https://doi.org/10.1007/BF00967965· OSTI ID:5654077

Childers, S R

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 (/sup 3/H)ATP as substrate, and the principal compound contaminating the (/sup 3/H)cyclic AMP peak, adenosine, is removed by incubation of assay tubes with small amounts of adenosine deaminase. The HPLC elution utilizes high resolution (3 microns) 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.

• Research Organization: Univ. of Florida College of Medicine, Gainesville
• OSTI ID: 5654077
• Journal Information: Neurochem. Res.; (United States), Journal Name: Neurochem. Res.; (United States) Vol. 2; ISSN NERED
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADENOSINE
AMIDINASES
AMP
ANIMALS
ATP
BODY
BRAIN
CENTRAL NERVOUS SYSTEM
CHROMATOGRAPHY
CYCLASES
ENZYMES
HYDROLASES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
LYASES
MAMMALS
NERVOUS SYSTEM
NON-PEPTIDE C-N HYDROLASES
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANS
RATS
RIBOSIDES
RODENTS
SEPARATION PROCESSES
SUBSTRATES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES