C Colling, R Gilles, M Cramer, N Nass, R Moka, L Jaenicke
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引用次数: 0
摘要
建立了一种基于单克隆抗体的快速、灵敏的3’:5’环AMP放射免疫测定方法。用蛋白偶联的2'- o -琥珀酰-3':5'-环AMP免疫小鼠，该单克隆抗体分别以琥珀酰cAMP (125I)碘酪氨酸甲酯和(3H) cAMP为示踪剂检测0.1和1摩尔cAMP。在毫摩尔范围内与其他腺苷核苷酸无交叉反应性;cGMP只有在过量500倍时才会干扰。直接分析植物和动物组织样品以及腺苷酸环化酶活性，或在有干扰物质的情况下进行适当的纯化。抗体结合法在各种组织中测定的环AMP水平与荧光乙烯基- camp高效液相色谱法测定的水平相一致。
Measurement of 3':5'cyclic AMP in biological samples using a specific monoclonal antibody.
A fast and sensitive radioimmunoassay for 3':5'cyclic AMP based on a monoclonal antibody has been worked out. Mice were immunized with protein-conjugated 2'-O-succinyl-3':5'-cyclic AMP. The monoclonal antibody detects 0.1 and 1 pmole cAMP with succinyl cAMP (125I)iodotyrosine methyl ester and (3H) cAMP, respectively, as tracers. It shows no cross-reactivity to other adenosine nucleotides up to the millimolar range; cGMP interferes only if present at a 500 fold excess. Plant and animal tissue samples as well as adenylate cyclase activity were analysed directly or after appropriate purification in case of interfering substances. Cyclic AMP levels measured in various tissues by the antibody binding assay correspond to those obtained by HPLC determination using fluorescent etheno-cAMP.
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