ArticleCold Spring Harbor protocols2012
Preparation of filamentous actin for polarized total internal reflection fluorescence microscopy (polTIRFM) motility assays.
Article in Cold Spring Harbor protocols, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed, 5 citations in OpenAlex.
- The polarized total internal reflection fluorescence microscopy (polTIRFM) twirling filament assay.Cold Spring Harbor protocols · 2012Article
- The polarized total internal reflection fluorescence microscopy (polTIRFM) processive motility assay for myosin V.Cold Spring Harbor protocols · 2012Article
- Fluorescent labeling of calmodulin with bifunctional rhodamine.Cold Spring Harbor protocols · 2012Article
- Orientation and rotational motions of single molecules by polarized total internal reflection fluorescence microscopy (polTIRFM).Cold Spring Harbor protocols · 2012Article
- Fluorescent labeling of myosin V for polarized total internal reflection fluorescence microscopy (polTIRFM) motility assays.Cold Spring Harbor protocols · 2012Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Polarized total internal reflection fluorescence microscopy (polTIRFM) can be used to detect the spatial orientation and rotational dynamics of single molecules. polTIRFM determines the three-dimensional angular orientation and the extent of wobble of a fluorescent probe bound to the macromolecule of interest. In this protocol, filamentous actin (F-actin) is polymerized from purified, monomeric actin (G-actin) for use in polTIRFM motility assays in which actin interacts with myosin. The procedures include (1) the preparation of unlabeled F-actin from G-actin; (2) the preparation of F-actin that is sparsely labeled with 6'-IATR (6'-iodoacetamidotetramethylrhodamine); and (3) the preparation of F-actin with a combination of unlabeled, biotinylated, and rhodamine-labeled monomers. Rhodamine-phalloidin actin, also used in polTIRFM assays, can be prepared using a procedure similar to the one for unlabeled actin.
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Registered trials
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