Using siRNA Silencing to Analyze ADAM17 in Macrophages

Markham, Matthew and Troeberg, Linda ORCID: https://orcid.org/0000-0003-0939-4651 (2023) Using siRNA Silencing to Analyze ADAM17 in Macrophages. In: Proteases and Cancer: Methods and Protocols. Methods in Molecular Biology (1). Springer, pp. 119-129. ISBN 978-1-0716-3588-9

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Abstract

Silencing expression with short interfering RNA (siRNA) is a rapid and cost-effective way to analyze the involvement of target genes in a range of biological processes. Here we describe isolation of primary human monocytes from peripheral blood and their in vitro differentiation to macrophages, followed by electroporation with siRNA to silence expression of a disintegrin and metalloproteinase 17 (ADAM17). This enables evaluation of ADAM17’s role in cleaving transmembrane proteins, such as its prototypic substrate tumor necrosis factor (TNF), by enzyme-linked immunosorbent assay (ELISA), flow cytometry, or immunoblotting.

Item Type:	Book Section
Faculty \ School:	Faculty of Medicine and Health Sciences > Norwich Medical School
Depositing User:	LivePure Connector
Date Deposited:	20 Aug 2024 11:30
Last Modified:	25 Sep 2024 10:45
URI:	https://ueaeprints.uea.ac.uk/id/eprint/96272
DOI:	10.1007/978-1-0716-3589-6_10

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