In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells

Ahmad Sharanek; Laura Raco; Vahab D. Soleimani; Arezu Jahani-Asl

STAR Protocols (Sep 2022)

In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells

Ahmad Sharanek,
Laura Raco,
Vahab D. Soleimani,
Arezu Jahani-Asl

Affiliations

Ahmad Sharanek: Department of Cellular and Molecular Medicine and University of Ottawa Brain and Mind Research Institute, University of Ottawa, 451 Smyth Road, Ottawa, ON K1H 8M5, Canada; Lady Davis Institute for Medical Research, Jewish General Hospital, Chemin de la Côte-Sainte-Catherine, Montréal, QC H3T 1E2, Canada
Laura Raco: Lady Davis Institute for Medical Research, Jewish General Hospital, Chemin de la Côte-Sainte-Catherine, Montréal, QC H3T 1E2, Canada; Division of Experimental Medicine, McGill University, 1001 Decarie Boulevard, Montreal, QC H4A 3J1, Canada
Vahab D. Soleimani: Lady Davis Institute for Medical Research, Jewish General Hospital, Chemin de la Côte-Sainte-Catherine, Montréal, QC H3T 1E2, Canada; Division of Experimental Medicine, McGill University, 1001 Decarie Boulevard, Montreal, QC H4A 3J1, Canada; Department of Human Genetics, McGill University, 3640 Rue University, Montréal, QC H3A OC7, Canada
Arezu Jahani-Asl: Department of Cellular and Molecular Medicine and University of Ottawa Brain and Mind Research Institute, University of Ottawa, 451 Smyth Road, Ottawa, ON K1H 8M5, Canada; Division of Experimental Medicine, McGill University, 1001 Decarie Boulevard, Montreal, QC H4A 3J1, Canada; Corresponding author

Journal volume & issue: Vol. 3, no. 3
p. 101554

Abstract

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Summary: Improper or aberrant protein-protein interactions can lead to severe human diseases including cancer. Here, we describe an adapted proximity ligation assay (PLA) protocol for the assessment of galectin-1-HOXA5 interaction in brain tumor stem cells (BTSCs). We detail the steps for culturing and preparation of BTSCs followed by PLA and detection of protein interactions in situ using fluorescent microscopy. This PLA protocol is optimized specifically for BTSCs and includes key controls for effective result analysis.For complete details on the use and execution of this protocol, please refer to Sharanek et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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