Streptavidin Acceptor Beads
Streptavidin Acceptor Beads
Product Details
Product Details
Product Specification
| Stability & Storage | Store at 2～8℃ away from light; product shelf life is 12 months. |
Background
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), 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, 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 protein interaction.
This product features a simple operation process, requires no washing, and offers high speed and sensitivity, enabling the detection of weak interactions.
Protocol
[Reagents Required]
Name |
Catalog Number |
| Protein A Donor Beads | UA086105 |
| Streptavidin Acceptor Beads | UA086090 |
| Universal Buffer 1 | UA086113 |
[Reference for the Assay Procedure]
Assay procedure |
Assay procedure 1 (37℃ rapid assay) |
Assay procedure 2 (room temperature assay) |
Step one: |
4 μL Tag1-M1 + 4 μL Tag2-M2 + 6 μL Donor Beads,Protect from light/green light |
4 μL Tag1-M1 + 4 μL Tag2-M2 + 6 μL Donor Beads,Protect from light/green light |
Incubation |
37℃ shaking incubation for 20 minutes,Protect from light/green light | Room temperature incubation for 60 minutes,Protect from light/green light |
Step two: |
Add 6 μL Acceptor Beads,Protect from light/green light |
Add 6 μL Acceptor Beads,Protect from light/Green light |
Incubation |
Incubate at 37℃ with shaking for 10 minutes,Protect from light/Green light |
Incubate at room temperature for 30 minutes,Protect from light/Green light |
ReadingInstrument reading |
Instrument reading |
Instrument reading |
[Performance Verification]
•Sample Preparation:
Use Universal Buffer 1 to pre-dilute biotinylated rabbit IgG (Bio-rIgG) to 15 μg/mL (100 nM) as the stock solution, then perform gradient dilutions according to the following scheme:
Number |
Final Concentration (nM) |
Universal Buffer 1 Volume (μL) |
High-Concentration Addition Volume (μL) |
C12 |
1.0E+01 |
210 |
90 μL of stock solution |
C11 |
3.0E+00 |
210 |
90 μL of C12 |
C10 |
1.0E+00 |
180 |
90 μL of 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 |
/ |
•Preparation of test reagents:
Name |
Preparation concentration |
Diluent |
| Protein A Donor Beads | 25 μg/mL |
Universal Buffer 1 |
| Streptavidin Acceptor Beads | 25 μg/mL |
Universal Buffer 1 |
•Results of the 37℃ incubation mode:
Highest signal: 389267
Lowest signal: 829
EC50 = 0.035 nM
•Results of room-temperature incubation:
Highest signal: 185868
Lowest signal: 407
EC50 = 0.029 nM
Guidelines
1. This experiment is light-sensitive. Perform all procedures, including preparation, dispensing, and incubation, under green light (illuminance below 100 LUX) to avoid light exposure.
2. This product is compatible with multi-mode 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 accompanying dilution buffer provided by our company 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 dispensing.
