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采用1,2-二氧杂环丁烷酶底物的化学发光报告基因检测法。

Chemiluminescent reporter gene assays with 1,2-dioxetane enzyme substrates.

作者信息

Olesen C E, Martin C S, Mosier J, Liu B, Voyta J C, Bronstein I

机构信息

Tropix, Inc., Bedford, Massachusetts 01730, USA.

出版信息

Methods Enzymol. 2000;305:428-50. doi: 10.1016/s0076-6879(00)05505-1.

Abstract

1,2-Dioxetane chemiluminescent substrates provide highly sensitive, quantitative detection with simple, rapid assay formats for the detection of reporter enzymes that are widely used in gene expression studies. Chemiluminescent detection methodologies typically provide up to 100-1000x higher sensitivities than can be achieved with the corresponding fluorescent or colorimetric enzyme substrates. The varieties of 1,2-dioxetane substrates available provides assay versatility, allowing optimization of assay formats with the available instrumentation, and are ideal for use in gene expression assays performed in both biomedical and pharmaceutical research. These assays are amenable to automation with a broad range of instrumentation for high throughput compound screening.

摘要

1,2 - 二氧杂环丁烷化学发光底物可通过简单、快速的检测形式实现高灵敏度的定量检测,用于检测在基因表达研究中广泛使用的报告酶。化学发光检测方法通常比相应的荧光或比色酶底物灵敏度高100 - 1000倍。现有的1,2 - 二氧杂环丁烷底物种类多样,可实现检测的多功能性,能根据现有仪器优化检测形式,非常适合用于生物医学和药物研究中的基因表达检测。这些检测适用于使用多种仪器进行自动化操作,以实现高通量化合物筛选。

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