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- W1567477963 abstract "Publisher Summary In vitro assays were originally applied to the study of myosin function and subsequently adapted for study of microtubule-based motors. Large, multicomponent complexes referred to as molecular motors can be adsorbed to inert surfaces yet retain the ability to generate force and translocate or move along microtubules. The development of videomicroscopy led, in part, to the discovery of kinesin and cytoplasmic dynein, and in observations about the mechanism of flagellar dynein. In vitro microscopic assays offer many advantages including use of small amounts of purified proteins, rapid and precise assessment of motility parameters, and convenient changes in the experimental variables. The goal of this chapter is to provide a description of the methods used for in vitro assay of flagellar dyneins. In general, the assay involves videomicroscopic observation of microtubule translocation across a slide surface to which purified dynein or dynein subunits have been adsorbed. These assays have proven useful in the study of dynein derived from Tetrahymena cilia, sea urchin sperm flagella, Paramecium cilia, and Chlamydomonas flagella. Most assays described to date make use of taxol-stabilized microtubules assembled from purified tubulin from either bovine or porcine brain. The chapter details the process of purification involving both cycles of assembly and disassembly and purification by chromatography. Dark-field microscopy has been used coupled to a silicon-intensified target camera to record microtubule movement on videotape. Both upright and inverted microscopes have been utilized. The experiment has been described with data analysis done. The primary measurement is the velocity of microtubule translocation." @default.
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- W1567477963 date "1995-01-01" @default.
- W1567477963 modified "2023-09-27" @default.
- W1567477963 title "Chapter 37 Microscopic Assays of Flagellar Dynein Activity" @default.
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- W1567477963 doi "https://doi.org/10.1016/s0091-679x(08)60818-3" @default.
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