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在聚(ADP-核糖)聚合酶纯化过程中通过酶联免疫吸附测定法对其进行快速检测及纯化方法的改进

Rapid detection of poly(ADP-ribose) polymerase by enzyme-linked immunosorbent assay during its purification and improvement of its purification.

作者信息

Sallmann F R, Plancke Y D, Poirier G G

机构信息

Poly(ADP-ribose) metabolism group, CHUL Research Center, CHUQ, Ste-Foy, Québec, Canada.

出版信息

Mol Cell Biochem. 1998 Aug;185(1-2):199-203. doi: 10.1023/a:1006861015142.

Abstract

We report a new detection method for the purification of poly(ADP-ribose) polymerase (PARP). PARP purification generates many fractions in which PARP is usually detected by a time consuming activity assay. The development of a new method was also needed in order to decrease the utilization of radioactivity. This new method, based on an enzyme-linked immunosorbent assay (ELISA), is very rapid, sensitive, and avoids most radioactivity. Moreover, to illustrate this method, a new matrix was used, the Heparin Sepharose. This matrix was chosen for its affinity for the DNA binding proteins and because it allows the separation of whole PARP from its proteolytic fragments.

摘要

我们报道了一种用于纯化聚(ADP - 核糖)聚合酶(PARP)的新检测方法。PARP纯化会产生许多级分,其中PARP通常通过耗时的活性测定来检测。为了减少放射性的使用,还需要开发一种新方法。这种基于酶联免疫吸附测定(ELISA)的新方法非常快速、灵敏,并且避免了大部分放射性。此外,为了说明这种方法,使用了一种新的基质,即肝素琼脂糖。选择这种基质是因为它对DNA结合蛋白具有亲和力,并且它能够将完整的PARP与其蛋白水解片段分离。

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