Human IL-1β Kit (HICA)

Testing 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: the acceptor microspheres are conjugated with antibody 1 targeting the protein of interest, while the donor microspheres are conjugated with streptavidin. Additionally, biotin-labeled antibody 2 is used. 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, the 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, quantitative analysis of the target protein can be achieved. This method offers advantages such as simplicity, rapid reaction, and high sensitivity.

Note: The recommended plate for use is a microplate (384-well or96-well plate, white, shallow well)

Reagent R3 must be protected from light during use. Sampling and incubation are recommended to be performed under green light (<100 LUX).

Recalibration is required for each test. Each concentration point of the standard should be tested with at least duplicate wells, and the calculation should be performed using a four-parameter (weight 1/Y²) fitting method.

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 be consistent with the test samples, and it must be used within 2 hours after reconstitution.

Components from different reagent kit batches must not be mixed.