title: Attachment of anti-GFP antibodies to microspheres for optical trapping experiments.
abstract: In vitro motility assays enabled the analysis of coupling between ATP hydrolysis and movement of myosin along actin filaments or kinesin along microtubules. Single-molecule assays using laser trapping have been used to obtain more detailed information about kinesins, myosins, and processive DNA enzymes. The combination of in vitro motility assays with laser-trap measurements has revealed detailed dynamic structural changes associated with the ATPase cycle. This protocol describes a method for attaching anti-GFP (green fluorescent protein) antibodies to microspheres. GFP-motor fusion proteins can then be adsorbed to the microspheres for use in single-molecule motility studies and optical trapping experiments.
authors: James A
authors: JA
authors: Spudich
authors: Sarah E
authors: SE
authors: Rice
authors: Ronald S
authors: RS
authors: Rock
authors: Thomas J
authors: TJ
authors: Purcell
authors: Hans M
authors: HM
authors: Warrick
mesh_terms: D000906:Antibodies / Q000276:immunology* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000276:immunology* / Q000378:metabolism*; D008863:Microspheres; D020409:Molecular Motor Proteins / Q000235:genetics / Q000378:metabolism; D052898:Optical Tweezers; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D000906:Antibodies; D020409:Molecular Motor Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins