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
Here is the English translation of the provided content, preserving the original formatting:```html
【Required Reagents】
Name |
Catalog Number |
| Protein A Donor Beads | UA086105 |
| Streptavidin Acceptor Beads | UA086090 |
| Universal Buffer 1 | UA086113 |
【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 Donor Beads,Light-protected/Green light |
4μL Tag1-M1 +4μL Tag2-M2+ 6μL Donor 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 Acceptor Beads,Light-protected/Green light |
Add 6μL Acceptor 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 |
Readout |
Instrument readout |
Instrument readout |
【Performance Validation】
•Sample Preparation:
Biotinylated rabbit IgG (Bio-rIgG) was pre-diluted to 15μg/mL (100nM) as stock solution using Universal Buffer 1, followed by serial dilution as below:
ID |
Final Concentration (nM) |
Universal Buffer 1 Volume (μL) |
High Concentration Addition Volume (μL) |
C12 |
1.0E+01 |
210 |
90μL 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 |
90μL C6 |
C4 |
1.0E-03 |
180 |
90μL C5 |
C3极> |
3.0E-04 |
210 |
90μL C4 |
C2 |
1.0E-04 |
180 |
90μL C3 |
C1 |
0 |
180 |
/ |
•Detection Reagent Preparation:
Name |
Preparation Concentration |
Diluent |
| Protein A Donor Beads | 25 μg/mL |
Universal Buffer 1 |
| Streptavidin Acceptor Beads | 25 μg/mL |
Universal Buffer 1 |
•37°C Incubation Mode Results:
Maximum signal: 389267
Minimum signal: 829
EC50= 0.035 nM
•Room Temperature Incubation Mode Results:
Maximum signal: 185868
Minimum 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.
