Anti-Mouse IgG Acceptor Beads
Anti-Mouse IgG Acceptor Beads
Product Details
Product Details
Product Specification
| Stability & Storage | Store away from light 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 and luminescence between donor 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 200 nm. Upon excitation at 680 nm, the donor beads generate singlet oxygen, which diffuses to the acceptor beads. The acceptor beads then undergo a redox reaction, emitting light at 615 nm. The signal intensity is directly proportional to the strength of the protein interaction.
This product offers a simple operation process, requires no washing, and provides fast results with high sensitivity, enabling the detection of 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. |
| Anti-Mouse IgG Acceptor Beads | UA086094 |
| Streptavidin Donor Beads | UA086104 |
| Universal Buffer 1 | UA086113 |
[Assay Procedure for Reference]
Assay Procedure |
Protocol 1 (37℃ Rapid Detection) |
Protocol 2 (Room Temperature Detection) |
Step 1: |
4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads,Avoid light/Green light |
4μL Tag1-M1 +4μL Tag2-M2+ 6μL Acceptor Beads,Avoid light/Green light |
Incubation |
Incubate with shaking at 37℃ for 20 minutes,Avoid light/Green light | Incubate at room temperature for 60 minutes,Avoid light/Green light |
Step 2: |
Add 6μL Donor Beads,Avoid light/Green light |
Add 6μL Donor Beads,Avoid light/Green light |
Incubation |
Incubate with shaking at 37℃ for 10 minutes,Avoid light/Green light |
Incubate at room temperature for 30 minutes,Avoid light/Green light |
Reading |
Instrument Reading |
Instrument Reading |
[Performance Validation]
•Sample Preparation:
Pre-dilute biotinylated mouse IgG (Bio-mIgG) to 15μg/mL (100nM) using Universal Buffer 1 as the stock solution, then perform serial dilutions according to the following scheme:
No. |
Final Concentration (nM) |
Universal Buffer 1 Volume (μL) |
High Concentration Add 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 |
3.0E-04 |
210 |
90μL C4 |
C2 |
1.0E-04 |
180 |
90μL C3 |
C1 |
0 |
180 |
/ |
•Detection Reagent Preparation:
Name |
Preparation Concentration |
Diluent |
| Anti-Mouse IgG Acceptor Beads | 25 μg/mL |
Universal Buffer 1 |
| Streptavidin Donor Beads | 25 μg/mL |
Universal Buffer 1 |
•Results for 37℃ Incubation Mode:
Maximum Signal: 5869294
Minimum Signal: 863
EC50= 0.137 nM
•Results for Room Temperature Incubation Mode:
Maximum Signal: 2281873
Minimum Signal: 446
EC50= 0.131 nM
Guidelines
1. This experiment is light-sensitive. Avoid exposure to light 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 multifunctional microplate readers equipped with Alpha detection modules.
3. Vortex thoroughly before use. Alternatively, briefly centrifuge (2000×g, 5–10 seconds) to ensure complete usage.
4. It is recommended to use the accompanying dilution buffer from our company for reagent preparation and sample dilution. If additional components are required, they can be directly added to this dilution buffer.
5. To ensure comparability of experimental data across different batches, strictly control incubation temperature and time.
6. Avoid generating bubbles during sample loading.
