Human IFN-γ Kit (HICA)

Test Principle:

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

The detection system includes 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 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 microsphere, triggering chemiluminescence. Conversely, if the target protein is absent, the microspheres remain too far apart, 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: The recommended plates are microplates (384 or 96 wells, white, shallow wells).

Reagent R3 must be protected from light. It is recommended to perform sample addition and incubation under green light (<100 LUX).

Each test requires recalibration. At least duplicate wells should be set for each concentration of the standard, and a four-parameter (weighting 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 sample matrix. Reconstituted calibrators should be used within 2 hours.

Components from different reagent kit lots must not be mixed.