Quantification of mitochondrial proteins in cultured cells by immuno-flow cytometry.
Immuno-flow cytometry was tested as a tool to estimate the cellular concentration of mitochondrial proteins in cultured cells, using cytochrome c oxidase as a model enzyme. Cells labelled with antibodies against cytochrome c oxidase, in which the amount of the enzyme was reduced by various extents, showed a linear relationship between the size of the signal obtained by immuno-flow cytometry and the amount of the enzyme. The determination by immuno-flow cytometry resulted in data comparable to the results obtained by immunoprecipitation and activity measurements. Since immuno-flow cytometry requires only limited numbers of cells, the method could especially be of value for diagnostic purposes. This is illustrated by the results obtained by comparing activity measurements and immuno-flow cytometry in the initial screening of cell lines derived from patients with deficiencies in the activity of cytochrome c oxidase.
Van den Bogert
C
C
E.C. Slater Institute for Biochemical Research, University of Amsterdam, The Netherlands.
Pennings
A
A
Dekker
H L
HL
Luciaková
K
K
Boezeman
J B
JB
Sinjorgo
K M
KM
D002478:Cells, Cultured; D030401:Cytochrome-c Oxidase Deficiency; D004318:Doxycycline / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000032:analysis*; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007106:Immune Sera; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology*; D011233:Precipitin Tests; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D007106:Immune Sera; D003576:Electron Transport Complex IV; D004318:Doxycycline