Protein L 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 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 can detect weak interactions.

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

[Required Reagents]

Name	Catalog No.
Protein L Acceptor Beads	UA086098
Streptavidin Donor Beads	UA086104
Universal Buffer 1	UA086113

[Detection Protocol for Reference]

Detection Protocol	Detection Protocol 1 (37℃Rapid Detection)	Detection Protocol 2 (Room Temperature Detection)
Step 1:	4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads，Avoid Light / Green Light	4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads，Avoid Light / Green Light
Incubation	37℃ Shaking Incubation 20 minutes，Avoid Light / Green Light	Room Temperature Incubation 60 minutes,,Avoid Light / Green Light
Step 2:	Add 6μL Donor Beads，Avoid Light / Green Light	Add 6μL Donor Beads，Avoid Light / Green Light
Incubation	37℃ Shaking Incubation 10 minutes，Avoid Light / Green Light	Room Temperature Incubation 30 minutes,Avoid Light / Green Light
Reading	Instrument Reading	Instrument Reading

[Performance Validation]

•Sample Preparation:

Use Universal Buffer 1 to pre-dilute Biotinylated Human IgG (Bio-hIgG) to 15μg/mL (100nM) as stock solution, then perform serial dilution 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	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
Protein L Acceptor Beads	25 μg/mL	Universal Buffer 1
Streptavidin Donor Beads	25 μg/mL	Universal Buffer 1

•37℃Incubation Mode Results:

Maximum Signal: 5917193

Minimum Signal: 1725

EC50= 0.402 nM

•Room Temperature Incubation Mode Results:

Maximum Signal: 2253290

Minimum Signal: 882

EC50= 0.371 nM

1. This experiment is light-sensitive; ensure all operations are performed in the dark. It is recommended to conduct preparation, sample loading, and incubation under green light (illuminance below 100 LUX). 2. This product is compatible with multi-mode 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 accompanying 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 loading.