Mouse IL-6 Kit (HICA)

Test Principle:

This kit employs a homogeneous immuno chemiluminescence assay (HICA) 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 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, 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.

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Note: Recommended to use microplates (384 or 96 well plates, white, shallow wells)

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

Recalibration is required for each test. At least duplicate wells should be set for each concentration of the standard, and a four-parameter (weighted 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 for calibrators should be consistent with the test samples. Reconstituted calibrators must be used within 2 hours.

Components from different reagent kit lots must not be mixed.