By Otto Z. Sellinger, Julio M. Azcurra (auth.), Neville Marks, Richard Rodnight (eds.)
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1966; Chappell and Hansford, 1969), rapid isolation procedures are often preferred. 39 40 Chapter 2 In isolating brain mitochondria, therefore, the method of choice will depend on the object of the research. For instance, oxidative phosphorylation and respiratory control require a method of isolating mitochondria as quickly as possible; location of enzymes forming part of or bound to mitochondria can only be achieved with preparations having minimal contamination. It is therefore impossible to give in this chapter a single method which will meet all these and other requirements.
The pellet is resuspended in 40 ml of the same medium and centrifuged at 5000 x g for 10 min. After the supernatant is decanted, the loosely packed white material on the top of the pellet is removed by shaking several times with small volumes of the medium. This follows the procedure of 10bsis (1963). The remaining tightly packed brown pellet represents the first mitochondrial fraction, F l , which is resuspended in a small volume of the medium. The supernatant plus the washing solutions is centrifuged at 5000 x g for 10 min after careful mixing.
Internat. Soc. Neurochem. (Milan), pp. 76--77. , and Gavendo, S. (\972) Cryobiology 9,192-197. Blomstrand, C. (1971) Thesis, University of Goteborg, Sweden. , and Hamberger, A. (\969)J. Neurochem. 16,1401-1407. , and Hamberger, A. (1970)J. Neurochem. 17, 1187-1195. Breckenridge, W. , and Morgan, I. G. (1972) FEBS Letters 22, 253-256. Cotman, C. W. (1972) In Research Methods in Neurochemistry, Vol. 1 (N. Marks and R. ), Plenum Press, New York, pp. 45-93. , and Rodriguez de Lores Arnaiz, G. (1969) In Handbook oj Neurochemistry, Vol.