Using siRNA Silencing to Analyze ADAM17 in Macrophages

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

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.
Original languageEnglish
Title of host publicationProteases and Cancer: Methods and Protocols
EditorsSalvatore Santamaria
PublisherSpringer
Pages119-129
Edition1
ISBN (Electronic)978-1-0716-3589-6
ISBN (Print)978-1-0716-3588-9, 978-1-0716-3591-9
DOIs
Publication statusPublished - 2 Dec 2023

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
ISSN (Print)1064-3745

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