RAT-LIVER ALCOHOL DEHYDROGENASE (RLADH)

Publisher Summary This chapter elaborates about rat-liver alcohol dehydrogenase (RLADH). RLADH has not been previously prepared in a pure state because of its instability. The stabilizing effects of several reagents were investigated and changes in the relative amounts of the isoenzymes and in the enzymatic activity were noticed. In the 35%–75% ammonium sulphate fraction twice as many enzyme units/kg of liver were found as previously reported. The activity of RLADH decreased with time at each stage of purification, even under optimal conditions. Some known stabilizing reagents like DTE, ME, and ethanol were added to the enzyme to prevent loss of activity. DTE was found to be very convenient, because the steroid activity could be estimated without dialysis even in relatively high concentrations of DTE. The purification of RLADH was checked by starch gel electrophoresis. Four separate bands (isoenzymes) were detected by enzyme activity staining. All the enzyme bands were on the cathodic side of the origin.