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- W4206827308 abstract "Filamentous actin (F-actin) organization within cells is regulated by a large number of actin-binding proteins that control actin nucleation, growth, cross-linking and/or disassembly. This protocol describes a technique – the actin co-sedimentation, or pelleting, assay – to determine whether a protein or protein domain binds F-actin and to measure the affinity of the interaction (i.e., the dissociation equilibrium constant). In this technique, a protein of interest is first incubated with F-actin in solution. Then, differential centrifugation is used to sediment the actin filaments, and the pelleted material is analyzed by SDS-PAGE. If the protein of interest binds F-actin, it will co-sediment with the actin filaments. The products of the binding reaction (i.e., F-actin and the protein of interest) can be quantified to determine the affinity of the interaction. The actin pelleting assay is a straightforward technique for determining if a protein of interest binds F-actin and for assessing how changes to that protein, such as ligand binding, affect its interaction with F-actin." @default.
- W4206827308 created "2022-01-25" @default.
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- W4206827308 date "2017-05-18" @default.
- W4206827308 modified "2023-09-30" @default.
- W4206827308 title "Measuring Protein Binding to F-actin by Co-sedimentation" @default.
- W4206827308 doi "https://doi.org/10.3791/55613-v" @default.
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