The basic principle and operation steps of human troponin T (Tn-T) chemiluminescence immunoassay kit

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Fundamental

This kit uses a double antibody sandwich method. The anti-Human Tn-T antibody was coated on the microtiter plate, and the Tn-T in the specimen or standard was bound to the coated antibody during the experiment, and the free component was washed away. Biotinylated anti-Human Tn-T antibody and horseradish peroxidase-labeled avidin were sequentially added. The anti-Human Tn-T antibody binds to Human Tn-T bound to the coated antibody, and biotin binds to avidin to form an immune complex, and the free component is washed away. The luminescent substrate mixture is added, the luminescent substrate is fluoresced under the catalysis of horseradish peroxidase, and the chemiluminescence value (CL) is measured by a chemiluminescence immunoassay, and the Tn-T concentration is positively correlated with the chemiluminescence value. The concentration of Tn-T in the specimen was determined by plotting a standard curve.

Steps

1. Add 100 μL of standard or sample to each well and incubate at 37 ° C for 90 minutes.

2. Pour the liquid in the well, pat dry, add 100 μL of biotinylated antibody working solution, incubate at 37 ° C for 60 minutes.

3. Wash 3 times

4. Add 100 μL of enzyme conjugate working solution and incubate at 37 ° C for 30 minutes.

5. Wash 5 times

6. Add 100 μL of substrate solution and incubate for 5 minutes at 37 °C

7. Readings

8. Calculation of results

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