Human IL-17F Kit (HICA)

Principle of the Assay:

This kit employs a homogeneous immuno chemiluminescence assay (HICA) based on the double-antibody sandwich principle for the detection of cytokine concentrations. The operation is simple and requires no washing steps.

The detection system consists of two types of microspheres: acceptor microspheres conjugated with antibody 1 targeting the protein of interest, and donor microspheres conjugated with streptavidin, along with biotin-labeled antibody 2. During the reaction, the target protein binds with the antibodies and microspheres to form an immune complex, bringing the two types of microspheres into close proximity. When the distance between the donor and acceptor microspheres is less than 200 nm, singlet oxygen generated by light excitation can transfer to the acceptor microspheres and trigger chemiluminescence. Conversely, if the target protein is absent, the distance between the microspheres remains too large, resulting in no signal.

By measuring the intensity of the chemiluminescent signal, quantitative analysis of the target protein can be achieved. This method offers advantages such as simplicity, rapid reaction, and high sensitivity.

Note: Microplates are recommended for use (384or96-well plates, white, shallow wells)

Reagent R3 must be protected from light. Sample addition and incubation should be performed under green light (<100 LUX).

Recalibration is required for each test. Each concentration point of the standard should be tested in at least duplicate, and a four-parameter (weight 1/Y²) fitting calculation should be applied.

Temperature and time must be controlled during incubation. The microplate should be covered with a film, and a microplate reader with ALPHA function is recommended.

The dilution matrix of the calibrator should match the test samples. Reconstituted calibrators must be used within 2 hours.

Components from different reagent kit lots must not be mixed.