Protocol for the production and purification of an i-Motif-specific nanobody
Summary: Intercalated motifs or i-Motifs (iMs) are nucleic acid structures formed by cytosine-rich sequences, which may regulate cellular processes and have broad applications in nanotechnology due to their pH-dependent nature. We have developed an iM-specific nanobody (iMbody) that can recognize iM...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723006962 |
| _version_ | 1797506346207150080 |
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| author | Mahdi Zeraati Samuel E. Ross Behnaz Aghaei Alvaro González Rajal Cecile King Marcel E. Dinger |
| author_facet | Mahdi Zeraati Samuel E. Ross Behnaz Aghaei Alvaro González Rajal Cecile King Marcel E. Dinger |
| author_sort | Mahdi Zeraati |
| collection | DOAJ |
| description | Summary: Intercalated motifs or i-Motifs (iMs) are nucleic acid structures formed by cytosine-rich sequences, which may regulate cellular processes and have broad applications in nanotechnology due to their pH-dependent nature. We have developed an iM-specific nanobody (iMbody) that can recognize iM DNA structures regardless of their sequences, making it a versatile research tool for studying iMs in various contexts. Here, we provide a protocol for the bacterial expression and His-tag purification of iMbody. We then describe procedures for performing ELISA and immunostaining using iMbody. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-10T04:31:21Z |
| format | Article |
| id | doaj.art-f7398032010c49589dab5a1ca8c35193 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-10T04:31:21Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f7398032010c49589dab5a1ca8c351932023-11-23T04:29:07ZengElsevierSTAR Protocols2666-16672023-12-0144102729Protocol for the production and purification of an i-Motif-specific nanobodyMahdi Zeraati0Samuel E. Ross1Behnaz Aghaei2Alvaro González Rajal3Cecile King4Marcel E. Dinger5School of Life and Environmental Sciences, Faculty of Science, The University of Sydney, Sydney, NSW 2006, Australia; Corresponding authorSchool of Life and Environmental Sciences, Faculty of Science, The University of Sydney, Sydney, NSW 2006, AustraliaSchool of Biotechnology and Biomolecular Sciences, Faculty of Science, The University of New South Wales Sydney, Sydney, NSW 2052, AustraliaSchool of Biotechnology and Biomolecular Sciences, Faculty of Science, The University of New South Wales Sydney, Sydney, NSW 2052, AustraliaSchool of Biotechnology and Biomolecular Sciences, Faculty of Science, The University of New South Wales Sydney, Sydney, NSW 2052, AustraliaSchool of Life and Environmental Sciences, Faculty of Science, The University of Sydney, Sydney, NSW 2006, Australia; Corresponding authorSummary: Intercalated motifs or i-Motifs (iMs) are nucleic acid structures formed by cytosine-rich sequences, which may regulate cellular processes and have broad applications in nanotechnology due to their pH-dependent nature. We have developed an iM-specific nanobody (iMbody) that can recognize iM DNA structures regardless of their sequences, making it a versatile research tool for studying iMs in various contexts. Here, we provide a protocol for the bacterial expression and His-tag purification of iMbody. We then describe procedures for performing ELISA and immunostaining using iMbody. : 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/S2666166723006962GenomicsMicroscopyMolecular BiologyAntibodyMolecular/Chemical ProbesProtein Biochemistry |
| spellingShingle | Mahdi Zeraati Samuel E. Ross Behnaz Aghaei Alvaro González Rajal Cecile King Marcel E. Dinger Protocol for the production and purification of an i-Motif-specific nanobody STAR Protocols Genomics Microscopy Molecular Biology Antibody Molecular/Chemical Probes Protein Biochemistry |
| title | Protocol for the production and purification of an i-Motif-specific nanobody |
| title_full | Protocol for the production and purification of an i-Motif-specific nanobody |
| title_fullStr | Protocol for the production and purification of an i-Motif-specific nanobody |
| title_full_unstemmed | Protocol for the production and purification of an i-Motif-specific nanobody |
| title_short | Protocol for the production and purification of an i-Motif-specific nanobody |
| title_sort | protocol for the production and purification of an i motif specific nanobody |
| topic | Genomics Microscopy Molecular Biology Antibody Molecular/Chemical Probes Protein Biochemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723006962 |
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