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Chapters
- 00:05Title
- 00:39Cell Culture for Immunofluorescence Microscopy
- 01:32Preparation of Transfection Complex and Transfection of Cells
- 03:08Live Cell Imaging and Indirect Immunofluorescence
- 05:40Results: Identifying Kinesin Cargo via Fluorescence Microscopy
- 06:50Conclusion
Here, a protocol is presented to identify Kinesin-1 cargos. A motorless mutant of the Kinesin-1 heavy chain (KIF5B) aggregates in the cytoplasm and induces aggregation of its cargos. Both aggregates are detected under fluorescence microscopy. A similar strategy can be employed to identify cargos of other motor proteins.
Tags
Kinesin-1CargosFluorescence MicroscopyMolecular MotorsDominant Negative KIF5B MutantsLive Cell ImagingIndirect Fluorescence ImagingImmunofluorescent StudiesCover GlassesCell CultureTransfection MediumTdTomato-tagged Wild Type KIF5B Expression PlasmidMotorless KIF5B Expression PlasmidCandidate Cargo Protein
