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ENZYME KINETICS IN A HISTOCHEMICAL SYSTEM

EARL P. BENDITT 1 and MARGARET ARASE 1

1 From the Department of Pathology of the University of Chicago, Chicago, Illinois and the University of Washington, Seattle, Washington

Modification of the Lineweaver and Burk method of handling the chemical kinetics of an enzyme system has been made to suit the histochemical conditions. Thus if the reaction is allowed to run to some fixed quantity of end point 1/v becomes proportion to the time necessary to reach this end point. In practice serial dilutions of substrate and serial increments in time of incubation are set up. Slides are then separated under the microscope into groups of equal intensity. A plot of 1/[S] v.s. t fits a straight line as expected. Competition between two substrates, one chromogenic the other non-chromogenic, can be demonstrated. One source of deviation from the theoretical due to spontaneous hydrolysis of substrate has been recognized and can be accounted for.


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