Anti-Rabbit IgG Acceptor Beads

Homogeneous Immuno Chemiluminescence Assay (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 high speed and sensitivity. It is capable of detecting weak interactions.

Specification	Fill Volume
250 μg	50 μL
5 mg	1 mL
25 mg	1 mL x 5

# Translation Result:```html

【Required Reagents】

Name	Catalog Number
Anti-Rabbit IgG Acceptor Beads	UA086095
Streptavidin Donor Beads	UA086104
Universal Buffer 1	UA086113

【Detection Protocol for Reference】

Detection Process	Detection Protocol 1 (37°C Rapid Detection)	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 minutes,Light-protected/Green light	​Room temperature incubation for 60 minutes,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 minutes,Light-protected/Green light	​Room temperature incubation for 30 minutes,Light-protected/Green light
Reading​	Instrument reading	Instrument reading

【Performance Validation】

•Sample Preparation:

Biotinylated rabbit IgG (Bio-rIgG) was pre-diluted to 15μg/mL (100nM) using Universal Buffer 1 as stock solution, then serially diluted according to the following scheme:

ID	Final Concentration (nM)	Universal Buffer 1 Volume (μL)	High Concentration Addition Volume (μL)
C12	1.0E+01	210	90μL stock solution
C11	3.0E+00	210	90μL C12
C10	1.0E+00	180	90μL C11
C9	3.0E-01	210	90μL C10
C8	1.0E-01	180	90μL C9
C7极e="border:1px solid #000000;text-align:center;vertical-align:middle;"> 3.0E-02	210	90μL C8
C6	1.0E-02	180	90μL C7
C5	3.0E-03	210	90μL C6
C4	1.0E-03	180	90μL C5
C3	3.0E-04	210	90μL C4
C2	1.0E-04	180	90μL C3
C1	0	180	/

•Detection Reagent Preparation:

Name	Preparation Concentration	Diluent
Anti-Rabbit IgG Acceptor Beads	25 μg/mL	Universal Buffer 1
Streptavidin Donor Beads	25 μg/mL	Universal Buffer 1

•37°C Incubation Mode Results:

Maximum signal: 4612453

Minimum signal: 646

EC50= 0.123 nM

•Room Temperature Incubation Mode Results:

Maximum signal: 1673088

Minimum signal: 305

EC50= 0.111 nM

```

1. This experiment is light-sensitive. Perform all procedures (preparation, sample loading, and incubation) under green light conditions (illuminance below 100 LUX) to avoid light exposure. 2. This product is compatible with multifunctional microplate readers equipped with an Alpha detection module. 3. Vortex thoroughly before use or briefly centrifuge (2000×g, 5–10 seconds) to ensure complete sample retrieval. 4. It is recommended to use the company's配套 (matching) diluent for reagent preparation and sample dilution. If additional components are required, they can be directly added to this diluent. 5. To ensure comparability of experimental data across different batches, strictly control incubation temperature and duration. 6. Avoid bubble formation during sample loading.