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Description
Human IgG Donor BeadsProduct Specification Stability & Storage Store in a dark place at 2 8C; 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), and acceptor beads recognize protein 2 (Tag2 label). When protein 1 binds to protein 2, the
Product Specification
| Stability & Storage | Store in a dark place at 2-8°C; 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), and acceptor beads recognize protein 2 (Tag2 label). When protein 1 binds to protein 2, the distance between the beads is less than 200 nm. Upon excitation at 680 nm, donor beads generate singlet oxygen, which diffuses to acceptor beads. The acceptor beads then undergo a redox reaction, emitting light at 615 nm. The signal intensity is proportional to the strength of protein interaction.
This product features a simple operation process, no washing steps, fast speed, high sensitivity, and the ability to detect weak interactions.
Protocol
[Required Reagents]
Name |
Catalog Number |
Human IgG Donor Beads |
UA086112 |
Streptavidin Acceptor Beads |
UA086090 |
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 hFC tag-M1 +4μL Biotin-M2+ 6μL Human IgG Donor Beads，Avoid Light / Green Light |
4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Human IgG Donor Beads，Avoid Light / Green Light |
Incubation |
37℃ Shaking Incubation 20 minutes，Avoid Light / Green Light |
Room Temperature Incubation 60 minutes,，Avoid Light / Green Light |
Step 2: |
Add 6μL Streptavidin Acceptor Beads，Avoid Light / Green Light |
Add 6μL Streptavidin Acceptor Beads，Avoid Light / Green Light |
Incubation |
37℃ Shaking Incubation 10 minutes，Avoid Light / Green Light |
Room Temperature Incubation 30 minutes，Avoid Light / Green Light |
Reading |
Instrument Reading |
Instrument Reading |
[Performance Validation]
•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 |
Human IgG Donor Beads |
25 μg/mL |
Universal Buffer 1 |
Streptavidin Acceptor Beads |
25 μg/mL |
Universal Buffer 1 |
•37℃ Incubation Mode Results:
Maximum Signal: 390419
Minimum Signal: 409
EC50= 0.076 nM
•Room Temperature Incubation Mode Results:
Maximum Signal: 194000
Minimum Signal: 181
EC50= 0.043 nM
Guidelines
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Exchange/Return Notes
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