一个简单和廉价的蛋白质结合试验环AMP在生物材料。

A. Geisler, R. Klysner, P. Thams, S. Christensen
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引用次数: 81

摘要

我们开发了一种简单廉价的测定生物材料中大容量cAMP的方法。该分析是基于3H-cAMP与兔骨骼肌分离蛋白的竞争性结合。在培养液中加入牛血清白蛋白以减少非特异性干扰。通过(NH4)2SO4沉淀进行自由和束缚放射性的分离,从而确定自由或束缚部分。在后一种情况下,所有程序都在同一管中进行,最后加入1.2 ml闪烁液进行计数。只有四分之一。该方法已应用于大鼠组织提取物和尿液中,具有良好的灵敏度、精密度和准确度。
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A simple and inexpensive protein binding assay for cyclic AMP in biological materials.
We have developed a simple and inexpensive method for largecapacity cAMP determination in biological materials. The assay is based on competitive binding of 3H-cAMP to proteins isolated from rabbit skeletal muscle. Bovine serum albumin is added to the incubate to reduce non-specific interference. Separation of free and bound radioactivity is performed by (NH4)2SO4 precipitation allowing determination of either free or bound fraction. In the latter case, all procedures are performed in the same tube, to which is finally added 1.2 ml of scintillation fluid for counting. By this only one fourth. The method has been applied to rat tissue extracts and urine with satisfactory sensitivity, precision and accuracy.
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