Clinical Chemistry · 1981 · 11 citations · 0 references
Life SciencesBiochemistryEndocrinologyAnalytical ChemistryHematology PhysiologyMetabolic StabilityIn Vitro Metabolic AssaysProtein DegradationRed Blood Cell PhysiologyHemolysis AssaysHemolysis MechanismsInsulin BiochemistryInsulin DegradationInsulin MeasurementOptimum TemperatureRed Blood Cell HemolysisTemperature Optimum
Abstract In vitro hemolysates of isolated human erythrocytes degrade 125I-labeled insulin. Ten- to 100-fold dilutions of the hemolysate give a proportionately decreased degradation of 125I-labeled insulin at 37 degrees C, while dilutions of up to eightfold do not. Like the control, diluted "Buffer G" containing 5 mmol/L Tris and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer alone, more than 500-fold dilutions of the hemolysate or boiled hemolysate (in buffer) caused negligible (less than 1%) degradation of the labeled insulin. We conclude that accurate insulin-binding data during erythrocyte insulin radioreceptor assay under optimum conditions (Clin. Chem. 23: 1590-1595, 1977) depend on avoiding or minimizing hemolysis.