Additional Information

Assay Principle

The human CDNF test is based on the quantitative sandwich enzyme immunoassay technique. Microtiter wells are pre-coated with human CDNF-specific monoclonal capture antibodies. Samples and standards are pipetted into microwells and human CDNF molecules present in the sample are bound by the capture antibodies. After incubation, unbound material is removed by washing the wells. Then, horseradish peroxidase (HRP) conjugated human CDNF-specific monoclonal detection antibodies bind to a different epitope of human CDNF molecules. After washing, the ready to use HRP substrate (TMB) is added to wells. The intensity of the colour produced is directly proportional to the amount of human CDNF in the sample. Colour development is then stopped by the addition of stop solution. Absorbance is measured at 450 nm.

Assay Procedure

1. Dilute 50 mL wash concentrate with 450 mL of distilled water.
2. Perform the dilution of each sample in the sample diluent (pink).
3. Add 100 μL samples and standards into appropriate wells in duplicate.
4. Incubate the covered microplate for 1 hr at RT on a microwell plate shaker (300 rpm).
5. Aspirate and wash the wells 4 times with 300 μL of washing solution.
6. Add 100 μL of enzyme conjugate (blue) into each well.
7. Incubate the covered microplate for 30 to 60 min at RT on a microwell plate shaker (300 rpm).
8. Aspirate and wash the wells 4 times with 300 μL of washing solution.
9. Add 100 μL of substrate solution into each well.
10. Incubate the covered microplate for 15 minutes at RT on a microwell plate shaker (300 rpm).
11. Stop the reaction by adding 50 μl of STOP solution into each well in the same order and time as for TMB distribution.
12. Read the absorbance at 450 nm immediately.

Typical Standard Curve