Related Experiment Videos
Rapid purification of cytochrome c oxidase from Paracoccus denitrificans
G C Steffens1, E Pascual, G Buse
1Institut für Biochemie, Rheinisch-Westfälische Technische Hochschule Aachen, Klinikum, Aachen, F.R.G.
Abstract:
Two methods are described for the purification of cytochrome c oxidase from Triton X-100 extracts of the periplasma membrane of Paracoccus denitrificans. The first is a large-scale procedure for the preparation of 100-250 nmol of cytochrome c oxidase (10-20 mg) in 1 week. The second is a rapid procedure for isolating up to 25 nmol in 2-3 days. Owing to the high yields given by fast protein liquid chromatography (FPLC) on Mono Q columns, the overall yield is about 20%, whereas the yield in many other previously published procedures does not exceed 10%. The use of FPLC on Mono Q also offers a considerable saving of time.