Anti-Human IgG Fc Donor Beads
Anti-Human IgG Fc Donor Beads
Product Details
Product Details
Product Specification
| Stability & Storage | Store in a dark place at 2-8℃; the product has a shelf life of 12 months. |
Background
Homogeneous Immuno Chemiluminescence Assay (HICA) is a homogeneous immunoassay method based on energy transfer and luminescence between donor beads and acceptor beads at close proximity.
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, no washing steps, fast speed, and high sensitivity, making it suitable for detecting weak interactions.
Components
Specification |
Quantity |
250 μg |
50 μL |
5 mg |
1 mL |
25 mg |
1 mL x 5 |
Protocol
[Required Reagents]
Name |
Catalog No. |
Anti-Human IgG Fc Donor Beads |
UA086110 |
Streptavidin Acceptor Beads |
UA086090 |
Universal Buffer 1 |
UA086113 |
[Reference Detection Protocol]
Procedure |
Protocol 1 (37℃ Rapid Assay) |
Protocol 2 (Room Temperature Assay) |
Step 1: |
4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Anti-Human IgG Fc Donor Beads，Protect from light/Green light |
4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Anti-Human IgG Fc 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 2: |
Add 6μL Streptavidin Acceptor Beads，Protect from light/Green light |
Add 6μL Streptavidin Acceptor Beads，Protect from light/Green light |
Incubation |
37℃ 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:
Dilute biotinylated human IgG (Bio-hIgG) to 15 μg/mL (100 nM) using Universal Buffer 1 as the stock solution, then perform serial dilutions according to the following scheme:
ID |
Final Concentration (nM) |
Universal Buffer 1 Volume (μL) |
High Concentration Add Volume (μL) |
C12 |
1.0E+01 |
210 |
90 μL Stock Solution |
C11 |
3.0E+00 |
210 |
90 μL C12 |
C10 |
1.0E+00 |
180 |
90 μL C11 |
C9 |
3.0E-01 |
210 |
90 μL C10 |
C8 |
1.0E-01 |
180 |
90 μL C9 |
C7 |
3.0E-02 |
210 |
90 μL C8 |
C6 |
1.0E-02 |
180 |
90 μL C7 |
C5 |
3.0E-03 |
210 |
90 μL C6 |
C4 |
1.0E-03 |
180 |
90 μL C5 |
C3 |
3.0E-04 |
210 |
90 μL C4 |
C2 |
1.0E-04 |
180 |
90 μL C3 |
C1 |
0 |
180 |
/ |
•Detection Reagent Preparation:
Name |
Working Concentration |
Diluent |
Anti-Human IgG Fc Donor Beads |
25 μg/mL |
Universal Buffer 1 |
Streptavidin Acceptor Beads |
25 μg/mL |
Universal Buffer 1 |
•Results at 37℃ Incubation:
Maximum Signal: 40063
Minimum Signal: 350
EC50= 0.083 nM
•Results at Room Temperature Incubation:
Maximum Signal: 27570
Minimum Signal: 172
EC50= 0.039 nM
Guidelines
1. This experiment is light-sensitive. Perform all procedures (e.g., preparation, loading, and incubation) under green light conditions (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 for reagent preparation and sample dilution. Additional components can be directly added to this buffer if required.
5. To ensure comparability of experimental data across batches, strictly control incubation temperature and duration.
6. Avoid bubble formation during loading.
