dos Santos, A. and Toseland, C. orcid.org/0000-0002-1641-7535 (2022) Magnetic tweezers in a microplate format. Journal of Visualized Experiments, 180. e62994. ISSN 1940-087X
Abstract
Mechanobiology describes how the physical forces and mechanical properties of biological material contribute to physiology and disease. Typically, these approaches are limited single-molecule methods, which restricts their availability. To address this need, a microplate assay was developed that enables mechanical manipulation while performing standard biochemical assays. This is achieved using magnets incorporated into a microplate lid to create multiple magnetic tweezers. In this format, force is exerted across biomolecules connected to paramagnetic beads, equivalent to a typical magnetic tweezer. The study demonstrates the application of this tool with FRET-based assays to monitor protein conformations. However, this approach is widely applicable to different biological systems ranging from measuring enzymatic activity through to the activation of signaling pathways in live cells.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2021 MyJoVE Corporation. This is an author-produced version of a paper subsequently published in Journal of Visualized Experiments. This version is distributed under the terms of the Creative Commons Attribution-NonCommercial Licence (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. You may not use the material for commercial purposes. |
Keywords: | mechanobiology; tweezers; 3D-printed; magnets; FRET; ensemble |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > Department of Human Metabolism (Sheffield) The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > Department of Oncology (Sheffield) |
Funding Information: | Funder Grant number CANCER RESEARCH UK C63579/A26206 MEDICAL RESEARCH COUNCIL MR/M020606/2 ENGINEERING AND PHYSICAL SCIENCE RESEARCH COUNCIL EP/V520469/1 |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 23 Sep 2021 10:27 |
Last Modified: | 09 Aug 2022 00:15 |
Status: | Published |
Publisher: | MyJove Corporation |
Refereed: | Yes |
Identification Number: | 10.3791/62994 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:178244 |