Anti-His Tag Acceptor Beads
Anti-His Tag Acceptor Beads
Product Details
Product Details
Product Specification
| Stability & Storage | Store away from light at 2-8℃; 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 for 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, no washing steps, fast speed, and high sensitivity, enabling the detection of weak binding.
Components
Specification |
Volume |
250 μg |
50 μL |
5 mg |
1 mL |
25 mg |
1 mL x 5 |
Protocol
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【Required Reagents】
Name |
Catalog Number |
| Anti-His Tag Acceptor Beads | UA086100 |
| 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,Protect from light/green light |
4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads,Protect from light/green light |
Incubation |
37°C shaking incubation for 20 minutes,Protect from light/green light | Room temperature incubation for 60 minutes,Protect from light/green light |
Step 2: |
Add 6μL Donor Beads,Protect from light/green light |
Add 6μL Donor Beads,Protect from light/green light |
Incubation |
37°C shaking incubation for 10 minutes,Protect from light/green light |
Room temperature incubation for 30 minutes,Protect from light/green light |
Reading |
Instrument reading |
Instrument reading |
【Performance Validation】
•Sample Preparation:
Biotinylated 6xHis (Bio-HH-6) was pre-diluted to 11μg/mL (10μM) 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+03 |
210 |
90μL stock |
C11 |
3.0E+02 |
210 |
90μL C12 |
C10 |
1.0E+02 |
180 |
90μL C11 |
C9 |
3.0E-01 |
210 |
90μL C10 |
C8 |
1.0E-01 |
180 |
90μL C9 |
C7 |
3.0E-00 |
210 |
90μL C8 |
C6 |
1.0E-00 |
180 |
90μL C7 |
C5 |
3.0E-01 |
210 |
90μL C6 |
C4 |
1.0E-01 |
180 |
90μL C5 |
C3 |
3.0E-02 |
210 |
90μL C4 |
C2 |
1.0E-02 |
180 |
90μL C3 |
C1 |
0 |
180 |
/ |
•Detection Reagent Preparation:
Name |
Preparation Concentration |
Diluent |
| Anti-His Tag Acceptor Beads | 25 μg/mL |
Universal Buffer 1 |
| Streptavidin Donor Beads | 25 μg/mL |
Universal Buffer 1 |
•37°C Incubation Mode Results:
Maximum signal: 222589
Minimum signal: 569
EC50= 3.919 nM
•Room Temperature Incubation Mode Results:
Maximum signal: 119656
Minimum signal: 316
EC50= 3.656 nM
```Guidelines
1. This experiment is light-sensitive. Perform all procedures under light-protected conditions. It is recommended to conduct preparation, sample loading, and incubation steps under green light (illuminance below 100 LUX).
2. This product is compatible with multifunctional microplate readers equipped with Alpha detection modules.
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. For special additive requirements, corresponding components 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.
