Human IL-2R Kit (HICA)

Principle of the Assay:

This kit employs a homogeneous immuno chemiluminescence assay (HICA) based on the dual-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: receptor 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 receptor microspheres is less than 200 nm, singlet oxygen generated by light excitation can transfer to the receptor microspheres and trigger 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, quantitative analysis of the target protein can be achieved. This method is characterized by its simplicity, rapid reaction, and high sensitivity.

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Note: It is recommended to use microplates (384 or 96-well plates, white, shallow wells) for the assay.

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 (weighting 1/Y²) curve fitting should be applied for calculation.

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

The dilution matrix for calibrators should be consistent with the sample matrix. Reconstituted calibrators must be used within 2 hours.

Components from different reagent kit lots must not be mixed.