Protein G Donor Beads
Protein G Donor Beads
Product Details
Product Details
Product Specification
| Stability & Storage | Store at 2-8°C protected 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, resulting in luminescence.
Donor beads recognize Protein 1 (Tag1 label), and 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 the protein interaction.
This product features a simple operation process, requires no washing, and offers high speed and sensitivity. It can detect weak binding 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 G Donor Beads |
UA086106 |
Streptavidin Acceptor Beads |
UA086090 |
Universal Buffer 1 |
UA086113 |
[Reference Detection Protocol]
Detection Protocol |
Protocol 1 (37℃ Rapid Detection) |
Protocol 2 (Room Temperature Detection) |
Step 1: |
4μL FC tag-M1 +4μL Biotin-M2+ 6μL Protein G Donor Beads, protect from light/green light |
4μL FC tag-M1 +4μL Biotin-M2+ 6μL Protein G Donor Beads, protect from light/green light |
Incubation |
Incubate at 37℃ with shaking for 20 minutes, protect from light/green light |
Incubate at room temperature 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 |
Incubate at 37℃ with shaking for 10 minutes, protect from light/green light |
Incubate at room temperature for 30 minutes, protect from light/green light |
Reading |
Instrument Reading |
Instrument Reading |
[Performance Validation]
•Sample Preparation:
Dilute biotinylated rabbit IgG (Bio-rIgG) to 15 μg/mL (100nM) using Universal Buffer1 as the stock solution, then perform serial dilutions according to the following scheme:
No. |
Final Concentration (nM) |
Volume of Universal Buffer 1 (μL) |
Volume of High Conc. Addition (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 |
/ |
•Detection Reagent Preparation:
Name |
Preparation Concentration |
Diluent |
Protein G Donor Beads |
25 μg/mL |
Universal Buffer 1 |
Streptavidin Acceptor Beads |
25 μg/mL |
Universal Buffer 1 |
•37℃ Incubation Mode Results:
Maximum Signal:933627
Minimum Signal:674
EC50= 0.045 nM
•Room Temperature Incubation Mode Results:
Maximum Signal:434224
Minimum Signal:329
EC50= 0.037 nM
Guidelines
1. This experiment is light-sensitive; take care to avoid light during operation. It is recommended to perform the preparation, sample addition, and incubation steps under green light conditions (illuminance below 100 LUX).
2. This product is compatible with multi-mode microplate readers equipped with an Alpha detection module.
3. Before use, vortex thoroughly to mix; a brief centrifugation (2000×g for 5–10 seconds) may also be performed to ensure complete recovery of the reagent.
4. We recommend using our company's dedicated dilution buffer for reagent preparation and analyte dilution; if special additives are required, they can be directly added to this dilution buffer.
5. To ensure comparability of experimental data across different batches, please strictly control the incubation temperature and time.
6. When adding samples, be careful to avoid introducing air bubbles.
