Human IL-10 Kit (HICA)

Test Principle:

This kit utilizes a homogeneous immuno chemiluminescence assay (HICA) based on the double-antibody sandwich method for the detection of cytokine concentrations. The operation is simple and requires no washing steps.

The detection system includes 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 by light excitation can transfer to the acceptor microspheres and trigger chemiluminescence. Conversely, if the target protein is absent, the distance between the microspheres remains too large, 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 simple operation, rapid reaction, and high sensitivity.

Name	Specification	Component Specification
ReagentR1A	500T	1mL/vial×1
	2000T	4ml/vial×1
	10000T	20ml/vial×1
ReagentR1F	500T	1mL/vial×1
	2000T	4ml/vial×1
	10000T	20ml/vial×1
ReagentR2	500T	1mL/vial×1
	2000T	4ml/vial×1
	10000T	20ml/vial×1
ReagentR3	500T	5mL/vial×1
	2000T	20ml/vial×1
	10000T	100ml/vial×1
Standard	500T	0.015μglyophilized powder×1
	2000T	0.015μglyophilized powder×2
	10000T	0.015μglyophilized powder×5
Standard Buffer	500T	6mL/vial×1
	2000T	12ml/vial×1
	10000T	30ml/vial×1

Note: Recommended plates are microplates (384or96-well plates, white, shallow well)

Reagent R3 must be protected from light. Sample addition and incubation are recommended to be performed under green light (<100 LUX). Each test requires recalibration. A minimum of 2 replicate wells should be set for each concentration point of the standard, and the calculation should be performed using a four-parameter (weighting: 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 match that of the test samples. Reconstituted calibrators must be used within 2 hours. Components from different reagent kit lots must not be mixed.