Anti-GST Tag 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 to generate luminescence.

Donor beads recognize protein 1 (Tag1 label), and 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. It is capable of detecting weak interactions.

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

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【Required Reagents】

Name	Catalog No.
Anti-GST Tag Acceptor Beads	UA086102
Streptavidin Donor Beads	UA086104
Universal Buffer 1	UA086113

 

【Detection Procedure (For Reference)】

Detection Steps	Procedure 1 (37°C Rapid Detection)	Procedure 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 shaking incubation 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 shaking incubation for 10 minutes,Light-protected/Green light	​Room temperature incubation for 30 minutes,Light-protected/Green light
Readout​	Instrument readout	Instrument readout

 

【Performance Validation】

•Sample Preparation:

Biotinylated GST (Bio-GST) was pre-diluted to 8.1μg/mL (300nM) using Universal Buffer 1 as the stock solution, followed by serial dilution as below:

ID	Final Concentration (nM)	Universal Buffer 1 Volume (μL)	High Concentration Addition Volume (μL)
C12	1.0E+01	270	30μL stock
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极p>	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-GST Tag Acceptor Beads	25 μg/mL	Universal Buffer 1
Streptavidin Donor Beads	25 μg/mL	Universal Buffer 1

 

 

•37°C Incubation Mode Results:

 

Max signal: 1142982

Min signal: 644

EC50= 0.8679nM

•Room Temperature Incubation Mode Results:

Max signal: 438266

Min signal: 295

EC50= 0.8027nM

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1. This experiment is light-sensitive; ensure all operations are performed in the dark. It is recommended to carry out preparation, sample loading, 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 retrieval. 4. It is recommended to use the company's配套 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 generating bubbles during sample loading.