Double-label immunoelectron microscopy for studying the colocalization of proteins in cultured cells.
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.
Hagiwara
Haruo
H
Department of Occupational Therapy, School of Health Sciences, Faculty of Medicine, Gunma University, Gunma, Japan.
Aoki
Takeo
T
Suzuki
Takeshi
T
Takata
Kuniaki
K
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
D000906:Antibodies; D000941:Antigens; C495540:CAV1 protein, human; D051242:Caveolin 1; D017979:Gold Colloid; D005780:Gelatin; D020696:rab5 GTP-Binding Proteins