In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722004348 |
| _version_ | 1818048316956475392 |
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| author | Ahmad Sharanek Laura Raco Vahab D. Soleimani Arezu Jahani-Asl |
| author_facet | Ahmad Sharanek Laura Raco Vahab D. Soleimani Arezu Jahani-Asl |
| author_sort | Ahmad Sharanek |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-10T10:19:45Z |
| format | Article |
| id | doaj.art-f1dd9ac7a9a64cc4a93d808488c184a4 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-10T10:19:45Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f1dd9ac7a9a64cc4a93d808488c184a42022-12-22T01:52:54ZengElsevierSTAR Protocols2666-16672022-09-0133101554In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cellsAhmad Sharanek0Laura Raco1Vahab D. Soleimani2Arezu Jahani-Asl3Department 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, CanadaLady 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, CanadaLady 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, CanadaDepartment 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 authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166722004348Cell BiologyCancerMicroscopyNeuroscienceMolecular/Chemical ProbesProtein Biochemistry |
| spellingShingle | Ahmad Sharanek Laura Raco Vahab D. Soleimani Arezu Jahani-Asl In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells STAR Protocols Cell Biology Cancer Microscopy Neuroscience Molecular/Chemical Probes Protein Biochemistry |
| title | In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells |
| title_full | In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells |
| title_fullStr | In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells |
| title_full_unstemmed | In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells |
| title_short | In situ detection of protein-protein interaction by proximity ligation assay in patient derived brain tumor stem cells |
| title_sort | in situ detection of protein protein interaction by proximity ligation assay in patient derived brain tumor stem cells |
| topic | Cell Biology Cancer Microscopy Neuroscience Molecular/Chemical Probes Protein Biochemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722004348 |
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