Stopped flow reaction rate measurement is the ideal tool for automation of enzymatic assays and for the study of enzyme inhibition.
In this example, spectrophotometric assay of glucose are biased the use of either dehydrogenase –NADH a reaction monitored at 337nm, or glucose oxidase quinoneimine dye monitored at 500nm.
The dehydrogenase assay of glucose is shown here as a series of superimposed reaction rate curves stopped flow period), obtained by injection of 20 microliters of glucose of increasing concentration. Since the volume of the flow cell is far smaller than the volume of the reacting mixture, it is essential to capture always the same section of the moving zone in the flow cell (Green arrow). This is achieved by selection of flow reversal volume.
Positioning Sample Zone in Flow Cell
2.2.10.
Hexokinase/glucose-6-phosphate dehydrogenase method (Sigma kit)
