Intracellular protein localization by immunoelectron microscopy.
Eukaryotic cells are characterized by the presence of a number of membrane-bound organelles which have differing degrees of internal structure. After synthesis in the cytoplasm, proteins must be targeted to the appropriate organelle and localized to the correct suborganellular compartment. We describe a method for immunoelectron microscopy that can be used to localize a protein not only to the correct organelle but to the appropriate suborganellular compartment. Cells are fixed to preserve subcellular structures and ultrathin sections are labeled with a monospecific antibody to the protein of interest. Protein-A gold is used to visualize the antigen-antibody complex by transmission electron microscopy allowing the intracellular location of the antigen to be determined. The methodology described was developed to study protein localization in Euglena but it is applicable to most organisms.
Osafune
Tetsuaki
T
Department of Life Sciences, Nippon Sport Science University, Yokohama, Japan.
Schwartzbach
Steven D
SD
D000818:Animals; D002736:Chloroplasts / Q000378:metabolism / Q000648:ultrastructure; D005055:Euglena / Q000378:metabolism / Q000648:ultrastructure*; D008565:Membrane Proteins / Q000378:metabolism / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron / Q000379:methods*; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure; D011506:Proteins / Q000378:metabolism / Q000648:ultrastructure*
D008565:Membrane Proteins; D011506:Proteins