Western blotting against tagged virulence determinants to study bacterial pathogenicity

Gili Aviv, Ohad Gal-Mor*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

2 Scopus citations

Abstract

Western blotting is a common approach to detect the presence of a target protein in biological samples or proteins mixture using specific antibodies. This method is also useful to study regulation of virulence determinants by analyzing changes in protein expression between different genetic backgrounds or under varying environmental conditions. To avoid the need to raise specific antibodies for each studied protein, commercial antibody against commonly used peptidic epitopes can be utilized if the right target tagged version is constructed. Here we describe a C-terminal fusion between a protein of interest and the two hemagglutinin A (2HA) tag. The tagged protein is cloned into a low-copy number vector and expressed under its native promoter in Salmonella enterica. Then, the expression of the tagged protein can be analyzed by Western blotting and commercially available anti-2HA antibodies.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages47-54
Number of pages8
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1734
ISSN (Print)1064-3745

Funding

FundersFunder number
German-Israeli Foundation for Scientific Research and Development1096.39.11/2010
Israel Science Foundation999/14
Ministry of Health, State of Israel3-0000-12435

    Keywords

    • Antibodies
    • Gel electrophoresis
    • Hemagglutinin
    • Immunoblotting
    • Protein tagging
    • Western blotting

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