Use of monoclonal antibodies in immuno-electron microscopy for the determination of subunit stoichiometry in oligomeric enzymes. There are three alpha-subunits in the F1-ATPase of Escherichia coli.
The subunit stoichiometry of oligomeric enzymes can be determined by immuno-electron microscopy using monoclonal antibodies against the individual subunits. Monoclonal antibodies against native F1-ATPase of Escherichia coli were prepared that were specific for the alpha-subunit. The immune complexes of F1 and monoclonal antibodies were isolated. Electron microscopy revealed the presence of three immunoglobulins per molecule of F1-ATPase. This unequivocally demonstrates an alpha 3 stoichiometry for the F1-ATPase of E. coli.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000302:isolation & purification; D002850:Chromatography, Gel; D004926:Escherichia coli / Q000201:enzymology*; D008854:Microscopy, Electron; D006180:Proton-Translocating ATPases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006180:Proton-Translocating ATPases