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使用改良的热位移分析评估线虫中受体-配体结合的方案。

Protocol to assess receptor-ligand binding in C. elegans using adapted thermal shift assays.

机构信息

Program in Innate Immunity, Division of Infectious Diseases and Immunology, Department of Medicine, UMass Chan Medical School, Worcester, MA 01655, USA.

Program in Innate Immunity, Division of Infectious Diseases and Immunology, Department of Medicine, UMass Chan Medical School, Worcester, MA 01655, USA.

出版信息

STAR Protoc. 2023 Sep 15;4(3):102477. doi: 10.1016/j.xpro.2023.102477. Epub 2023 Jul 31.

Abstract

The Caenorhabditis elegans genome encodes a greatly expanded number of nuclear hormone receptors, many of which remain orphaned. Here, we present a protocol to assess ligand-receptor binding in C. elegans using an adapted cellular thermal shift assay and isothermal dose response. We describe steps for growing C. elegans and preparing lysates and compounds. We also detail how to perform and quantify these assays. This protocol can be used to study any soluble receptor. For complete details on the use and execution of this protocol, please refer to Peterson et al. (2023)..

摘要

秀丽隐杆线虫基因组编码了大量扩展的核激素受体,其中许多仍然是孤儿受体。在这里,我们使用改良的细胞热移位分析和等温剂量反应方法,提供了一种评估秀丽隐杆线虫中配体-受体结合的方案。我们描述了培养秀丽隐杆线虫和制备溶胞产物及化合物的步骤。我们还详细介绍了如何进行和定量这些测定。该方案可用于研究任何可溶性受体。有关该方案使用和执行的完整详细信息,请参阅 Peterson 等人(2023 年)。

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