Abstract
In the case of the integral membrane protein Na+/K+-ATPase, preparation of highly concentrated samples for IR difference spectroscopy often leads to inactivation of the enzyme. Therefore, we compared the activity of Na+/K+-ATPase using different techniques of sample preparation. The loss of activity can be minimized by cooling the sample to 10 degrees C and by the addition of glycerol and dithiothreitol. The activity of Na+/K+-ATPase isolated from pig kidney is independent of the protein concentration whereas the enzyme from shark rectal gland is inactivated at concentrations above 1 microg/microL and is thus unsuitable for IR experiments.
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