Human IL-8 Kit (HICA)

Testing Principle:

This kit employs a homogeneous immuno chemiluminescence assay (HICA) based on the double-antibody sandwich method for detecting the concentration of cytokines. 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 upon light excitation can transfer to the acceptor microspheres, triggering chemiluminescence. Conversely, if the target protein is absent, the distance between the microspheres is too large, and no signal is produced.

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

Note: The recommended plate for use is a microplate (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).

Recalibration is required for each test. At least duplicate wells should be set for each concentration point of the standard, and a four-parameter (weight 1/Y²) fitting calculation should be used.

Temperature and time must be controlled during incubation. Microplates 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. Use within 2 hours after reconstitution.

Components from different reagent kit batches must not be mixed.