Glutathione Peroxidase (GSH-PX) Test Box Specification I. Experimental apparatus: test tube, micropipette, vortex mixer, centrifuge, 37°C constant temperature water bath, visible spectrophotometer (wavelength 412nm) . Second, the scope of application: The kit can measure animal serum, plasma, tissue and whole blood, red blood cells in the GSH-PX activity. Third, the determination of significance: glutathione peroxidase (GSH-PX) is an important peroxide decomposition of the body's extensive enzyme, which specifically catalyzes the reduction of glutathione (GSH) on hydrogen Reduction of peroxides. It is generally thought that it can eliminate harmful peroxide metabolites in the cell and block the lipid peroxidation chain-locking reaction, thereby protecting cell membrane structure and function integrity. This enzyme contains selenium, which is an integral part of GSH-PX. Each gram of enzyme contains 4 gram atoms of selenium. The active center of GSH-PX is selenocysteine. Measuring GSH-PX activity can be used as a biochemical measure of selenium levels in the body. Fourth, the operation table: 1, the enzymatic reaction: (reagent one application liquid in advance at 37oC pre-temperature) control tube 1mmol/L GSH (ml) to be tested hemolyzed (ml) reagent one application liquid reagent two (ml) to be tested dissolved Blood (ml) 37oC Water bath pre-warm 5 minutes 0.1 37oC Water bath accurate reaction 5 minutes 20.20.2 Determination tube 0.20.20.12 Mix well, 3500-4000 rpm, centrifuge for 10 minutes, take supernatant 1 ml for color reaction. 2. Color Reaction: Blank GSH Standard Solution Solvent (ml) 20 μmol/L GSH Standard (ml) Supernatant (ml) Reagent 3 Application Solution (ml) Reagent 4 Application Solution (ml) Reagent 5 Application Solution (ml)10.250.0510.250.0511110.250.05110.250.05 standard tube control tube tube mix, after standing at room temperature for 15 minutes, 412nm, 1cm optical diameter than the color cup, double distilled water, zero, measure the OD value of each tube. V. Calculation formula: 1. Calculation of GSH-PX activity in whole blood: Definition: Each 4μl of whole blood is required to react at 37°C for 5 minutes, and the effect of non-enzymatic reaction is deducted to reduce the concentration of GSH in the reaction system by 1μmol/L. An enzyme activity unit. 2. Calculation of the activity of GSH-PX in tissue: Definition: It is defined as every milligram of protein, and the effect of non-enzymatic reaction is deducted per minute, so that the concentration of GSH in the reaction system is reduced by 1 μmol/L as an enzyme activity unit. 3. Calculation of GSH-PX enzyme activity in serum: Definition: Each 0.1ml serum is allowed to react at 37°C for 5 minutes, and the non-enzymatic reaction is subtracted to reduce the GSH concentration in the reaction system by 1μmol/L as an enzyme activity unit.
December 21, 2021
