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使用针对三链体、聚(ADP-核糖)、DNA-RNA杂交体和Z-DNA的单克隆抗体检测异常核酸。

Detection of unusual nucleic acids with monoclonal antibodies specific for triplexes, poly(ADP-ribose), DNA-RNA hybrids, and Z-DNA.

作者信息

Lee J S, Latimer L J, Sibley J T

机构信息

Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.

出版信息

Anal Biochem. 1989 May 1;178(2):373-7. doi: 10.1016/0003-2697(89)90655-6.

Abstract

Monoclonal antibodies which are specific for several unusual nucleic acids are now available. These include Jel 318 which is specific for triplexes, ADP-1 specific for poly(ADP-ribose), Jel 99 specific for RNA-DNA duplexes, and Jel 150 specific for Z-DNA. With the aid of these antibodies and an immunoblotting procedure, unusual nucleic acids can be detected and the amount estimated from a variety of sources. The method involves binding the nucleic acid to either nitrocellulose or Zeta Probe (a cationic nylon membrane), probing with the appropriate monoclonal antibody, followed by addition of an 125I-labeled anti-mouse second antibody. The blot is then developed by autoradiography. The technique is extremely sensitive and can be used to estimate unusual nucleic acids from crude cell extracts.

摘要

现在已有针对几种特殊核酸的单克隆抗体。其中包括对三链体具有特异性的Jel 318、对聚(ADP - 核糖)具有特异性的ADP - 1、对RNA - DNA双链体具有特异性的Jel 99以及对Z - DNA具有特异性的Jel 150。借助这些抗体和免疫印迹程序,可以从各种来源检测特殊核酸并估计其含量。该方法包括将核酸与硝酸纤维素或Zeta Probe(一种阳离子尼龙膜)结合,用适当的单克隆抗体进行检测,然后加入125I标记的抗小鼠二抗。然后通过放射自显影使印迹显影。该技术极其灵敏,可用于从粗细胞提取物中估计特殊核酸的含量。

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