Human IL-12p70 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 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 types of microspheres into close proximity. When the distance between the donor and acceptor microspheres is less than 200 nm, singlet oxygen generated upon light excitation can transfer to the acceptor microspheres, 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 is characterized by its simplicity, rapid reaction, and high sensitivity.

Note: Recommended plates are microplates (384or96-well plates, white, flat-bottom)

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

Recalibration is required for each test. At least two replicates of each standard concentration should be tested, and a four-parameter (weighted 1/Y²) fitting calculation should be applied.

Temperature and time should be strictly 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 be consistent with the test samples. The reconstituted solution should be used within 2 hours.

Components from different reagent kit batches must not be mixed.