One-step immunoaffinity purification of complex I subunits from beef heart mitochondria.
Polypeptides of beef heart mitochondrial complex I were isolated from 15 mg of solubilized beef heart mitochondria using antibodies immobilized on an agarose chromatography column. The preparation was examined by SDS electrophoresis and Western blotting using affinity-purified antibodies to complex I and compared to beef heart complex I purified according to the conventional method of Hatefi and Rieske. There was a high degree of homology between the two preparations as judged by SDS-polyacrylamide electrophoresis and by immunoblotting with seven affinity-purified antibodies to various complex I subunits. This method could be applied to the preparation of complex I subunits from small samples such as human muscle biopsy specimens.
Haines
A M
AM
Department of Neuroscience, Royal Free Hospital School of Medicine, London, United Kingdom.
Cooper
J M
JM
Morgan-Hughes
J A
JA
Clark
J B
JB
Schapira
A H
AH
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D015151:Immunoblotting; D007163:Immunosorbent Techniques; D008929:Mitochondria, Heart / Q000201:enzymology*; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000096:biosynthesis* / Q000276:immunology
D016660:NAD(P)H Dehydrogenase (Quinone)