Protein A Donor Beads
Protein A Donor Beads
Product Details
Product Details
Product Specification
| Stability & Storage | Store in a dark place at 2-8°C; product shelf life is 12 months. |
Background
The Homogeneous Immuno Chemiluminescence Assay (HICA) is a homogeneous immunoassay method based on energy transfer luminescence between donor beads and acceptor beads.
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 directly proportional to the strength of protein interaction.
This product features a simple operation process, requires no washing, offers fast results, and high sensitivity. It is capable of detecting weak interactions.
Components
Specification |
Fill Volume |
250 μg |
50 μL |
5 mg |
1 mL |
25 mg |
1 mL x 5 |
Protocol
【Required Reagents】
Name |
Catalog No. |
Protein A Donor Beads |
UA086105 |
Streptavidin Acceptor Beads |
UA086090 |
Universal Buffer 1 |
UA086113 |
【Assay Protocol for Reference】
Assay Steps |
Protocol 1 (37℃ Rapid Assay) |
Protocol 2 (Room Temperature Assay) |
Step 1: |
4μL FC tag-M1 +4μL Biotin-M2+ 6μL Protein A Donor Beads，Protect from light / Green light |
4μL FC tag-M1 +4μL Biotin-M2+ 6μL Protein A Donor Beads，Protect from light / Green light |
Incubation |
Incubate at 37℃ with shaking for 20 min, Protect from light / Green light |
Incubate at room temperature for 60 min,，Protect from light / Green light |
Step 2: |
Add 6μL Streptavidin Acceptor Beads，Protect from light / Green light |
Add 6μL Streptavidin Acceptor Beads，Protect from light / Green light |
Incubation |
Incubate at 37℃ with shaking for 10 min，Protect from light / Green light |
Incubate at room temperature for 30 min，Protect from light / Green light |
Reading |
Instrument reading |
Instrument reading |
【Performance Validation】
•Sample Preparation:
Use Universal Buffer 1 to pre-dilute Biotinylated Rabbit IgG (Bio-rIgG) to 15μg/mL（100nM）as a stock solution, then perform serial dilutions according to the following scheme:
No. |
Final Concentration (nM) |
Universal Buffer 1 Volume (μL) |
Volume of Higher Concentration Added (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 |
/ |
•Assay Reagent Preparation:
Name |
Working Concentration |
Diluent |
Protein A Donor Beads |
25 μg/mL |
Universal Buffer 1 |
Streptavidin Acceptor Beads |
25 μg/mL |
Universal Buffer 1 |
•37℃ 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. Avoid light exposure during operation. It is recommended to perform preparation, sample loading, and incubation steps under green light (illuminance below 100 LUX).
2. This product is compatible with microplate readers equipped with Alpha detection modules.
3. Vortex thoroughly before use, or briefly centrifuge (2000×g, 5–10 seconds) to ensure complete usage.
4. It is recommended to use the 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 time.
6. Avoid generating bubbles during sample loading.
