Anti-Mouse IgG Donor Beads
Anti-Mouse 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, requiring no washing, with fast speed and high sensitivity. It can detect weak binding.
Components
Specification |
Fill Volume |
250 μg |
50 μL |
5 mg |
1 mL |
25 mg |
1 mL x 5 |
Protocol
【Required Reagents】
Name |
Catalog No. |
Anti-Mouse IgG Donor Beads |
UA086107 |
Streptavidin Acceptor Beads |
UA086090 |
Universal Buffer 1 |
UA086113 |
【Assay Protocol for Reference】
Assay Protocol |
Protocol 1 (37℃ Rapid Assay) |
Protocol 2 (Room Temperature Assay) |
Step 1: |
4μL mFC tag-M1 +4μL Biotin-M2+ 6μL Anti-Mouse IgG Donor Beads,Avoid light/Green light |
4μL mFC tag-M1 +4μL Biotin-M2+ 6μL Anti-Mouse IgG Donor Beads,Avoid light/Green light |
Incubation |
37℃ Incubate with shaking for 20 minutes,Avoid light/Green light |
Incubate at room temperature for 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℃ Incubate with shaking for 10 minutes,Avoid light/Green light |
Incubate at room temperature for 30 minutes,Avoid light/Green light |
Reading |
Instrument Reading |
Instrument Reading |
【Performance Validation】
•Sample Preparation:
Dilute biotinylated mouse IgG (Bio-mIgG) in Universal Buffer1 to a pre-dilution of 15μg/mL (100nM) as a stock solution, then perform serial dilutions according to the following scheme:
No. |
Final Concentration (nM) |
Universal Buffer 1 Volume (μL) |
High Concentration Add 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 |
Anti-Mouse IgG Donor Beads |
25 μg/mL |
Universal Buffer 1 |
Streptavidin Acceptor Beads |
25 μg/mL |
Universal Buffer 1 |
•37℃ Incubation Mode Results:
Maximum Signal:335611
Minimum Signal:320
EC50= 0.030 nM
•Room Temperature Incubation Mode Results:
Maximum Signal:135470
Minimum Signal:156
EC50= 0.021 nM
Guidelines
