Principle and procedure of human brain-derived neurotrophic factor (BDNF) chemiluminescence immunoassay kit

product description:

Fundamental:

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

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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