title: The use of streptavidin conjugates as immunoblot loading controls and mitochondrial markers for use with Dictyostelium discoideum.
abstract: The loading controls used for quantitative immunoblotting of mammalian proteins are not appropriate for use with Dictyostelium discoideum. Actin levels, for example, change greatly during Dictyostelium development. In addition, Dictyostelium-specific antibodies for other potential control proteins are not commercially available. Here we demonstrate the use of labeled streptavidin to detect biotinylated mitochondrial 3-methylcrotonyl-CoA carboxylase α (MCCC1), providing a robust and convenient tool for quantitative normalization of Dictyostelium Western blots, as well as fluorescently labeling mitochondria for microscopy of fixed cells.
authors.affiliation: Beatson Institute for Cancer Research, Glasgow, UK.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Davidson
authors.forename: Jason S
authors.initials: JS
authors.lastname: King
authors.forename: Robert H
authors.initials: RH
authors.lastname: Insall
mesh_terms: D015415:Biomarkers / Q000032:analysis; D019735:Carbon-Carbon Ligases / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D004023:Dictyostelium / Q000737:chemistry* / Q000276:immunology; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D015151:Immunoblotting / Q000592:standards*; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D019809:Streptavidin / Q000737:chemistry*
chemical_list: D015415:Biomarkers; D005456:Fluorescent Dyes; D024101:Mitochondrial Proteins; D019809:Streptavidin; D019735:Carbon-Carbon Ligases; C021573:methylcrotonoyl-CoA carboxylase