A highly sensitive adenylate cyclase assay

A highly sensitive adenylate cyclase assay method has been developed which employs sequential chromatography on columns of Dowex cation exchange resin and aluminum oxide. With the use of [α- 32P]ATP as substrate, this method permits the nearly complete separation of cyclic [ 32P]AMP formed from the...

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Published in	Analytical biochemistry Vol. 58; no. 2; pp. 541 - 548
Main Authors	Salomon, Yoram, Londos, Constantine, Rodbell, Martin
Format	Journal Article
Language	English
Published	United States Elsevier Inc 01.04.1974
Subjects	Adenylyl Cyclases - analysis Adenylyl Cyclases - metabolism Aluminum Animals Cell Membrane - enzymology Chromatography Chromatography, Ion Exchange Evaluation Studies as Topic Glucagon - pharmacology Kinetics Liver - cytology Liver - enzymology Methods Microchemistry Phosphorus Radioisotopes Rats Time Factors
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Abstract	A highly sensitive adenylate cyclase assay method has been developed which employs sequential chromatography on columns of Dowex cation exchange resin and aluminum oxide. With the use of [α- 32P]ATP as substrate, this method permits the nearly complete separation of cyclic [ 32P]AMP formed from the substrate and other 32P-containing compounds, i.e., 32P in the assay blanks was barely detectable. In comparative studies, this method was found to be considerably more sensitive than previously reported methods. The high sensitivity of this method permits detection of the small amounts of cyclic AMP formed at low enzyme concentrations or at early time points in kinetic studies.
AbstractList	A highly sensitive adenylate cyclase assay method has been developed which employs sequential chromatography on columns of Dowex cation exchange resin and aluminum oxide. With the use of [α- 32P]ATP as substrate, this method permits the nearly complete separation of cyclic [ 32P]AMP formed from the substrate and other 32P-containing compounds, i.e., 32P in the assay blanks was barely detectable. In comparative studies, this method was found to be considerably more sensitive than previously reported methods. The high sensitivity of this method permits detection of the small amounts of cyclic AMP formed at low enzyme concentrations or at early time points in kinetic studies.
Author	Londos, Constantine Rodbell, Martin Salomon, Yoram
Author_xml	– sequence: 1 givenname: Yoram surname: Salomon fullname: Salomon, Yoram – sequence: 2 givenname: Constantine surname: Londos fullname: Londos, Constantine – sequence: 3 givenname: Martin surname: Rodbell fullname: Rodbell, Martin
BackLink	https://www.ncbi.nlm.nih.gov/pubmed/4827395$$D View this record in MEDLINE/PubMed
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References	Lowry, Rosebrough, Farr, Randall (BIB6) 1951; 193 White, Zenser (BIB2) 1971; 41 Rodbell, Lin, Salomon (BIB9) 1974; 249 (1973) personal communication. Neville (BIB5) 1968; 154 Ramanchandran (BIB3) 1971; 43 Rodbell (BIB7) 1972; Vol. 3 Krishna, Weiss, Brodie (BIB1) 1968; 163 Pohl, Birnbaumer, Rodbell (BIB4) 1971; 246 Lowry (10.1016/0003-2697(74)90222-X_BIB6) 1951; 193 Rodbell (10.1016/0003-2697(74)90222-X_BIB9) 1974; 249 Ramanchandran (10.1016/0003-2697(74)90222-X_BIB3) 1971; 43 White (10.1016/0003-2697(74)90222-X_BIB2) 1971; 41 Pohl (10.1016/0003-2697(74)90222-X_BIB4) 1971; 246 10.1016/0003-2697(74)90222-X_BIB8 Krishna (10.1016/0003-2697(74)90222-X_BIB1) 1968; 163 Rodbell (10.1016/0003-2697(74)90222-X_BIB7) 1972; Vol. 3 Neville (10.1016/0003-2697(74)90222-X_BIB5) 1968; 154
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Snippet	A highly sensitive adenylate cyclase assay method has been developed which employs sequential chromatography on columns of Dowex cation exchange resin and...
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SubjectTerms	Adenylyl Cyclases - analysis Adenylyl Cyclases - metabolism Aluminum Animals Cell Membrane - enzymology Chromatography Chromatography, Ion Exchange Evaluation Studies as Topic Glucagon - pharmacology Kinetics Liver - cytology Liver - enzymology Methods Microchemistry Phosphorus Radioisotopes Rats Time Factors
Title	A highly sensitive adenylate cyclase assay
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