Nickel Chelate Acceptor Beads

Homogeneous Chemiluminescence Immunoassay (HICA) is a homogeneous immunoassay method based on energy transfer between donor beads and acceptor beads at close proximity, resulting in luminescence.

Donor beads recognize protein 1 (Tag1 label), while acceptor beads recognize protein 2 (Tag2 label). When protein 1 binds to protein 2, the distance between the beads becomes less than 200nm. Upon excitation at 680nm, the donor beads generate singlet oxygen, which diffuses to the acceptor beads. The acceptor beads then undergo a redox reaction, emitting light at 615nm. The signal intensity is directly proportional to the strength of the protein interaction.

This product features a simple operation process, requires no washing, and offers fast results with high sensitivity, enabling the detection of weak interactions.

【Required Reagents】

Name	Catalog Number
Nickel Chelate Acceptor Beads	UA086093
Streptavidin Donor Beads	UA086104
Universal Buffer 3	UA086114

 

【Detection Protocol for Reference】

Detection Steps	Protocol 1 (37°C Rapid Detection)	Protocol 2 (Room Temperature Detection)
​Step 1:	4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads,Light-protected/Green light	4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads,Light-protected/Green light
Incubation	​37°C with shaking for 20 min,Light-protected/Green light	​Room temperature for 60 min,Light-protected/Green light
​Step 2:	Add 6μL Donor Beads,Light-protected/Green light	Add 6μL Donor Beads,Light-protected/Green light
Incubation	​37°C with shaking for 10 min,Light-protected/Green light	​Room temperature for 30 min,Light-protected/Green light
Readout​	Instrument readout	Instrument readout

 

【Performance Validation】

•Sample Preparation:

His-tagged M protein (His-M1) was pre-diluted to 20μg/mL (1μM) using Universal Buffer 3 as the stock solution, followed by serial dilution as below:

ID	Final Concentration (nM)	Universal Buffer 3 Volume (μL)	High-Concentration Addition Volume (μL)
C12	5.00E+01	210	90μL stock
C11	1.50E+01	210	90μL C12
C10	5.00E+00	180	90μL C11
C9	1.50E+00	210	90μL C10
C8	5.00E-01	180	90μL C9
C7	1.50E-01	210	90μL C8
C6极>	5.00E-02	180	90μL C7
C5	1.50E-02	210	90μL C6
C4	5.00E-03	180	90μL C5
C3	1.50E-03	210	90μL C4
C2	5.00E-04	180	90μL C3
C1	0	180	/

 

•Detection Reagent Preparation:

Name	Preparation Concentration	Diluent
Nickel Chelate Acceptor Beads	25 μg/mL	Universal Buffer 3
Streptavidin Donor Beads	25 μg/mL	Universal Buffer 3

 

 

•37°C Incubation Mode Results:

 

Max signal: 1287443

Min signal: 1609

EC50= 1.423 nM

 

 

•Room Temperature Incubation Mode Results:

Max signal: 673976

Min signal: 1610

EC50= 0.785 nM

1. This experiment is light-sensitive; ensure all operations are performed in the dark. It is recommended to conduct preparation, sample addition, and incubation steps under green light (illuminance below 100 LUX). 2. This product is compatible with multifunctional microplate readers equipped with an Alpha detection module. 3. Vortex thoroughly before use. Alternatively, briefly centrifuge (2000×g, 5–10 seconds) to ensure complete sample retrieval. 4. It is recommended to use the company’s配套dilution buffer for reagent preparation and sample dilution. If additional components are required, they can be directly added to this buffer. 5. To ensure comparability of experimental data across different batches, strictly control incubation temperature and duration. 6. Avoid generating bubbles during sample addition.