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通过膜印迹法提高放射自显影和荧光自显影的灵敏度。

Increased sensitivity of autoradiography and fluorography by membrane blotting.

作者信息

Quéméneur E, Simonnet F

机构信息

CEA-DIEP, Gif-sur-Yvette, France.

出版信息

Biotechniques. 1995 Jan;18(1):100, 102-3.

PMID:7702834
Abstract

The detection of low amounts of 3H-, 14C- or 35S-labeled compounds can be difficult with x-ray films. Here we report the benefits of blotting labeled molecules that have been electrophoretically separated to a membrane prior to autoradiographic or fluorographic detection. Blotting allows a reduction, by a factor of 5-10, in exposure times compared with dried gels. When combined with a rapid and safe alternative to the impregnation of blots with liquid scintillant, a further 3-fold to 5-fold increase in sensitivity is achieved. Bands containing as low as 1 Bq 14C or 100 to 10(3) Bq 3H could be detected after 24 hours of exposure. This enhancement in sensitivity may lead to new applications of 3H and 14C.

摘要

用X光胶片检测少量的³H、¹⁴C或³⁵S标记化合物可能会有困难。在此我们报告,在进行放射自显影或荧光成像检测之前,将经电泳分离的标记分子印迹到膜上的好处。与干燥的凝胶相比,印迹可使曝光时间缩短5至10倍。当与一种快速且安全的替代方法(即用液体闪烁剂浸渍印迹)相结合时,灵敏度可进一步提高3至5倍。曝光24小时后,可检测到低至1 Bq的¹⁴C或100至10³ Bq的³H的条带。这种灵敏度的提高可能会带来³H和¹⁴C的新应用。

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