ABSTRACT
There are two principal steps are used to preparation of subcellular membranes: homogenization of the tissue or cells, followed by fractionation of the organelles and membranes. This fractionation process is such a vast and complex area that it is not possible to provide more than a few basic themes. Moreover, the recovery in isolated particles of functional activities known to be associated with those organelles in vivo attests to the validity of the procedure. Malate dehydrogenase and succinate dehydrogenase are the most commonly used markers. Malate dehydrogenase activity is measured by the oxidation of NADH in the presence of oxaloacetate, via the decrease in absorbance of the NADH at 340 nm. With the advent of new techniques in DNA and protein analysis, together with the availability of monoclonal antibodies to proteins and their ready detection by immunoblotting after separation on Polyacrylamide gels.
