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一种用于测量细胞内cAMP的微量放射免疫测定法。

A micro-radioimmunoassay for the measurement of intracellular cAMP.

作者信息

Berg W, Hornbeck P V

机构信息

Department of Medicine, University of Maryland Medical School, Baltimore 21201.

出版信息

Biotechniques. 1993 Jul;15(1):56-9.

PMID:7689853
Abstract

We have developed a rapid and sensitive micro-radioimmunoassay for the detection of cellular cAMP. The assay employs a simple method for preparing cellular lysates that requires as few as 500 cells per data point. Cellular lysates are incubated with cAMP-specific antibody complexes and [125I]cAMP in 96-well plates, harvested onto glass fiber filters and analyzed using a commercial autoradiograph imaging system. This method provides significant savings in reagents and may be particularly useful in situations where only a limited number of cells are available.

摘要

我们已经开发出一种用于检测细胞内环磷酸腺苷(cAMP)的快速灵敏的微量放射免疫测定法。该测定法采用一种制备细胞裂解物的简单方法,每个数据点仅需500个细胞。将细胞裂解物与cAMP特异性抗体复合物及[125I]cAMP在96孔板中孵育,收集到玻璃纤维滤器上,并用商用放射自显影片成像系统进行分析。这种方法可显著节省试剂,在细胞数量有限的情况下可能特别有用。

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