Human TNF-β Kit (HICA)

Test Principle:

This kit employs a homogeneous immuno chemiluminescence assay (HICA) for the detection of cytokine concentrations, featuring a simple operation process with no washing steps required.

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 upon light excitation can transfer to the receptor microsphere, 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: A microplate (384 or 96-well plate, white, shallow well) is recommended.

Reagent R3 should be protected from light during use, and sample addition and incubation are recommended to be performed under green light (<100 LUX) conditions.

Recalibration is required for each test, with at least duplicate wells for each concentration point of the standard. A four-parameter (weighted 1/Y²) curve fitting method should be used for calculation.

Temperature and time should be strictly 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, and the reconstituted calibrator should be used within 2 hours.

Components from different reagent kit lots must not be mixed.