Protein G Acceptor Beads
Protein G Acceptor 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 at close proximity to generate chemiluminescence.
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 proportional to the strength of the 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 |
Fill Volume |
250 μg |
50 μL |
5 mg |
1 mL |
25 mg |
1 mL x 5 |
Protocol
[Required Reagents]
Name |
Catalog Number |
| Protein G Acceptor Beads | UA086092 |
| Streptavidin Donor Beads | UA086104 |
| Universal Buffer 1 | UA086113 |
[Detection Procedure for Reference]
Detection Procedure |
Detection Procedure 1 (37℃Rapid Detection) |
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℃ Shaking incubation 20 minutes,Protect from light/Green light | Room temperature incubation 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℃ Shaking incubation 10 minutes,Protect from light/Green light |
Room temperature incubation 30 minutes,Protect from light/Green light |
Reading |
Instrument Reading |
Instrument Reading |
[Performance Verification]
•Sample Preparation:
Use Universal Buffer 1 to pre-dilute Biotinylated Rabbit IgG (Bio-rIgG) to 15μg/mL (100nM) as a stock solution, then perform gradient dilution according to the following scheme:
ID |
Final Concentration (nM) |
Universal Buffer 1 Volume (μL) |
High Concentration Addition Volume (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 Acceptor Beads | 25 μg/mL |
Universal Buffer 1 |
| Streptavidin Donor Beads | 25 μg/mL |
Universal Buffer 1 |
•37℃Incubation Mode Results:
Maximum Signal: 7087102
Minimum Signal: 707
EC50= 0.234 nM
•Room Temperature Incubation Mode Results:
Maximum Signal: 2604143
Minimum Signal: 386
EC50= 0.208 nM
Guidelines
1. This experiment is light-sensitive. Avoid exposure to light during operation. It is recommended to perform preparation, sample loading, and incubation under green light (illuminance below 100 LUX).
2. This product is compatible with microplate readers equipped with an Alpha detection module.
3. Vortex thoroughly before use, or 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 buffer.
5. To ensure comparability of experimental data across different batches, strictly control incubation temperature and time.
6. Avoid generating bubbles during sample loading.
