Solid-phase preparation of vimentin-type intermediate filaments for immunoassays.
A simple method is described for the preparation of vimentin-type intermediate filaments for solid-phase immunoassays. Bovine aortic endothelial cells were grown to confluency in multiwells, and extracted with Triton X-100 and a high-salt buffer. Residual protein in the wells consisted of 53% vimentin as judged by SDS electrophoresis, and the recovery of vimentin was 41%. Methanol-fixed and dried filaments can be used directly for immunological assays.
D000818:Animals; D002417:Cattle; D002458:Cell Fractionation / Q000379:methods; D003599:Cytoskeleton / Q000648:ultrastructure*; D004727:Endothelium / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D007118:Immunoassay / Q000379:methods; D007382:Intermediate Filaments / Q000276:immunology / Q000648:ultrastructure*; D014746:Vimentin / Q000276:immunology* / Q000302:isolation & purification
D014746:Vimentin