Product Details
Product Details
Product Specification
| Stability & Storage | Store in a dark place at 2~8℃; product shelf life is 12 months. |
Background
Homogeneous Immuno Chemiluminescence Assay (HICA) is a homogeneous immunoassay based on energy transfer between donor beads and acceptor beads at close proximity, resulting in luminescence.
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 the protein interaction.
This product offers a simple, wash-free operation process, with high speed and sensitivity, enabling the detection of weak interactions.

Components
Specification |
Volume |
250 μg |
50 μL |
5 mg |
1 mL |
25 mg |
1 mL x 5 |
Protocol
[Required Reagents]
Name |
Catalog No. |
| Anti-FITC Acceptor Beads | UA086101 |
| Streptavidin Donor Beads | UA086104 |
| Universal Buffer 1 | UA086113 |
[Detection Protocol for Reference]
Protocol |
Protocol 1 (Rapid detection at 37°C) |
Protocol 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 |
Shake incubate at 37°C for 20 minutes,Protect from light / Green light | Incubate at room temperature for 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 |
Shake incubate at 37°C 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 FITC-BSA-Biotin to 6.6 μg/mL (100 nM) as stock solution using Universal Buffer 1, then perform serial dilutions according to the following scheme:
No. |
Final Concentration (nM) |
Universal Buffer 1 Volume (μL) |
High Conc. Addition 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 |
Preparation Concentration |
Diluent |
| Anti-FITC Acceptor Beads | 25 μg/mL |
Universal Buffer 1 |
| Streptavidin Donor Beads | 25 μg/mL |
Universal Buffer 1 |
•Results for 37°C Incubation Mode:

Maximum Signal: 2779107
Minimum Signal: 1906
EC50= 0.290 nM
•Results for Room Temperature Incubation Mode:

Maximum Signal: 1095538
Minimum Signal: 754
EC50= 0.334 nM
Guidelines
