Purification and succinylation of cyclic GMP from large volume samples and radioimmunoassay of succinyl cyclic GMP. |
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| Authors: | T Asakawa T R Russell R Ho |
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| Affiliation: | Earl W. Sutherland Research Laboratories Department of Biochemistry, University of Miami School of Medicine Miami, Florida 33136 USA |
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| Abstract: | Improved procedures for isolation of cyclic GMP and cyclic AMP and radioimmunoassay of cyclic GMP with succinylation are described. Procedures involved include modified chromatography on alumina and succinylation of cyclic GMP followed by purification of succinyl cyclic GMP on a Dowex AG 1×8 column. These procedures are convenient and applicable to any volume up to 50 ml of tissue extracts and especially for isotonic incubation mixtures. This assay system is sensitive to 6 femtomoles of cyclic GMP/tube. On radioimmunoassay, free and antibody bound [125I]-labeled cyclic GMP are separated by Millipore filtration. Cyclic GMP levels in several tissue samples were determined in order to show the applicability of the procedures. |
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| Keywords: | cGMP (cyclic GMP) guanosine 3′,5′-monophosphate cAMP (cyclic AMP) adenosine 3′,5′-monophosphate ScGMP 2′-0-succinyl guanosine 3′,5′-monophosphate |
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