ArticleJournal of visualized experiments : JoVE2017
Measuring Protein Binding to F-actin by Co-sedimentation.
Article in Journal of visualized experiments : JoVE, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 17 citations in OpenAlex.
- IVNS1ABP mutation drives cellular senescence in newly identified progeroid neuropathy.Nature communications · 2026Article
- Cardiomyopathy-associated and basic residue mutations in myopalladin alter actin binding, bundling, and structural stability.Protein science : a publication of the Protein Society · 2026Article
- Carnosine enhances actomyosin-ATPase activity under acidic conditions: the role of carnosine in a rigor mortis.Journal of animal science · 2026Article
- Mapping the interaction surface between CaNature communications · 2025Article
- Spatial proteomics of ER tubules reveals CLMN, an ER-actin tether at focal adhesions that promotes cell migration.Cell reports · 2025Article
- Purification of modified mammalian actin isoforms for in vitro reconstitution assays.European journal of cell biology · 2023Review
- Article
- Induction of filopodia formation by α-Actinin-2 via RelA with a feedforward activation loop promoting overt bone marrow metastasis of gastric cancer.Journal of translational medicine · 2023Article
- LILAC: enhanced actin imaging with an optogenetic Lifeact.Nature methods · 2023Article
- Hyperglycemia and hyperlipidemia can induce morphophysiological changes in rat cardiac cell line.Biochemistry and biophysics reports · 2021Article
- Cardiomyopathy Mutation Alters End-to-End Junction of Tropomyosin and Reduces Calcium Sensitivity.Biophysical journal · 2020Article
- A Novel Actin Binding Drug withAntimicrobial agents and chemotherapy · 2019Article
- Structural basis of the filamin A actin-binding domain interaction with F-actin.Nature structural & molecular biology · 2018Article
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
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.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.