Anti-Human IgG Donor Beads
Anti-Human IgG Donor Beads
Product Details
Product Details
Product Specification
| Stability & Storage | Store at 2～8°C away 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, 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 features a simple operation process, requires no washing, and offers fast results with high sensitivity. It is capable of detecting weak interactions.
Components
Specification |
Volume |
250 μg |
50 μL |
5 mg |
1 mL |
25 mg |
1 mL x 5 |
Protocol
[Required Reagents]
Name |
Cat. No. |
Anti-Human IgG Donor Beads |
UA086109 |
Streptavidin Acceptor Beads |
UA086090 |
Universal Buffer 1 |
UA086113 |
[Assay Procedure for Reference]
Assay Procedure |
Assay Procedure 1 (37°C Rapid Assay) |
Assay Procedure 2 (Room Temperature Assay) |
Step 1: |
4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Anti-Human IgG Donor Beads, Protect from light / Green light |
4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Anti-Human IgG Donor Beads, Protect from light / Green light |
Incubation |
Shake and 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 Streptavidin Acceptor Beads, Protect from light / Green light |
Add 6μL Streptavidin Acceptor Beads, Protect from light / Green light |
Incubation |
Shake and incubate at 37°C for 10 minutes, Protect from light / Green light |
Incubate at room temperature for 30 minutes, Protect from light / Green light |
Readout |
Instrument Readout |
Instrument Readout |
[Performance Validation]
•Sample Preparation:
Pre-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:
No. |
Final Concentration (nM) |
Volume of Universal Buffer 1 (μL) |
Volume of High Conc. Addition (μL) |
C12 |
1.0E+01 |
210 |
90 μL 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 |
Working Concentration |
Diluent |
Anti-Human IgG Donor Beads |
25 μg/mL |
Universal Buffer 1 |
Streptavidin Acceptor Beads |
25 μg/mL |
Universal Buffer 1 |
•37°C Incubation Mode Results:
Maximum Signal:47550
Minimum Signal:192
EC50= 0.067 nM
•Results of Room Temperature Incubation Mode:
Maximum Signal:23402
Minimum Signal:92
EC50= 0.036 nM
Guidelines
