title: Double-label immunoelectron microscopy for studying the colocalization of proteins in cultured cells.
abstract: Multiple label immunoelectron microscopy localizes and detects multiple antigens in cells and tissues. In double labeling, two kinds of primary antibodies from different animal species are used after being mixed in a single solution. To distinguish the different antigens, secondary antibodies should be labeled with colloidal gold particles of different diameter. Generally, the secondary antibody that is used for detecting the antigen with lower distribution density is labeled with smaller-sized gold particles. In this chapter, double-label immunoelectron microscopy of gelatin-embedded cultured cells using the cryosectioning technique is described.
authors.affiliation: Department of Occupational Therapy, School of Health Sciences, Faculty of Medicine, Gunma University, Gunma, Japan.
authors.forename: Haruo
authors.initials: H
authors.lastname: Hagiwara
authors.forename: Takeo
authors.initials: T
authors.lastname: Aoki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Suzuki
authors.forename: Kuniaki
authors.initials: K
authors.lastname: Takata
mesh_terms: D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000941:Antigens / Q000032:analysis / Q000276:immunology; D051242:Caveolin 1 / Q000032:analysis / Q000276:immunology; D002478:Cells, Cultured; D016616:Cryoultramicrotomy; D042783:Endothelial Cells / Q000166:cytology / Q000276:immunology / Q000648:ultrastructure; D005780:Gelatin / Q000737:chemistry; D017979:Gold Colloid / Q000737:chemistry / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D016253:Microscopy, Immunoelectron / Q000379:methods*; D010316:Particle Size; D013194:Staining and Labeling / Q000379:methods*; D016610:Tissue Embedding; D020696:rab5 GTP-Binding Proteins / Q000032:analysis / Q000276:immunology
chemical_list: D000906:Antibodies; D000941:Antigens; C495540:CAV1 protein, human; D051242:Caveolin 1; D017979:Gold Colloid; D005780:Gelatin; D020696:rab5 GTP-Binding Proteins