Total Internal Reflection Fluorescence (TIRF) Single-Molecule Assay to Analyze the Motility of Kinesin
The motile parameters of kinesin superfamily proteins are fundamental to intracellular transport. Single-molecule motility assays using total internal reflection fluorescence (TIRF) microscopy are a gold standard technique for measuring the motile parameters of kinesin motors. With this technique, o...
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Bio-protocol LLC
2024-12-01
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| Series: | Bio-Protocol |
| Online Access: | https://bio-protocol.org/en/bpdetail?id=5135&type=0 |
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| _version_ | 1825206619830484992 |
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| author | Tomoki Kita Shinsuke Niwa |
| author_facet | Tomoki Kita Shinsuke Niwa |
| author_sort | Tomoki Kita |
| collection | DOAJ |
| description | The motile parameters of kinesin superfamily proteins are fundamental to intracellular transport. Single-molecule motility assays using total internal reflection fluorescence (TIRF) microscopy are a gold standard technique for measuring the motile parameters of kinesin motors. With this technique, one can evaluate the velocity, run length, and binding frequency of kinesins on microtubules by directly observing their motility. This protocol provides a comprehensive procedure for single molecule assays of kinesins, including the preparation of labeled microtubules, the measurement of kinesin motility via TIRF microscopy, and the quantification of kinesin motor parameters. |
| format | Article |
| id | doaj-art-4e93a8d0fe374acdb41fe14157b0e72b |
| institution | Kabale University |
| issn | 2331-8325 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Bio-protocol LLC |
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| series | Bio-Protocol |
| spelling | doaj-art-4e93a8d0fe374acdb41fe14157b0e72b2025-02-07T08:16:21ZengBio-protocol LLCBio-Protocol2331-83252024-12-01142410.21769/BioProtoc.5135Total Internal Reflection Fluorescence (TIRF) Single-Molecule Assay to Analyze the Motility of KinesinTomoki Kita0Shinsuke Niwa1Graduate School of Life Sciences, Tohoku University, Miyagi, JapanGraduate School of Life Sciences, Tohoku University, Miyagi, JapanFrontier Research Institute for Interdisciplinary Sciences (FRIS), Tohoku University, Miyagi, JapanThe motile parameters of kinesin superfamily proteins are fundamental to intracellular transport. Single-molecule motility assays using total internal reflection fluorescence (TIRF) microscopy are a gold standard technique for measuring the motile parameters of kinesin motors. With this technique, one can evaluate the velocity, run length, and binding frequency of kinesins on microtubules by directly observing their motility. This protocol provides a comprehensive procedure for single molecule assays of kinesins, including the preparation of labeled microtubules, the measurement of kinesin motility via TIRF microscopy, and the quantification of kinesin motor parameters.https://bio-protocol.org/en/bpdetail?id=5135&type=0 |
| spellingShingle | Tomoki Kita Shinsuke Niwa Total Internal Reflection Fluorescence (TIRF) Single-Molecule Assay to Analyze the Motility of Kinesin Bio-Protocol |
| title | Total Internal Reflection Fluorescence (TIRF) Single-Molecule Assay to Analyze the Motility of Kinesin |
| title_full | Total Internal Reflection Fluorescence (TIRF) Single-Molecule Assay to Analyze the Motility of Kinesin |
| title_fullStr | Total Internal Reflection Fluorescence (TIRF) Single-Molecule Assay to Analyze the Motility of Kinesin |
| title_full_unstemmed | Total Internal Reflection Fluorescence (TIRF) Single-Molecule Assay to Analyze the Motility of Kinesin |
| title_short | Total Internal Reflection Fluorescence (TIRF) Single-Molecule Assay to Analyze the Motility of Kinesin |
| title_sort | total internal reflection fluorescence tirf single molecule assay to analyze the motility of kinesin |
| url | https://bio-protocol.org/en/bpdetail?id=5135&type=0 |
| work_keys_str_mv | AT tomokikita totalinternalreflectionfluorescencetirfsinglemoleculeassaytoanalyzethemotilityofkinesin AT shinsukeniwa totalinternalreflectionfluorescencetirfsinglemoleculeassaytoanalyzethemotilityofkinesin |