Protein L Donor Beads
Protein L 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 in 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, and offers fast results with high sensitivity. It is capable of 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 L Donor Beads |
UA086111 |
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 Protein L Donor Beads，Protect from Light/Green Light |
4μL hFC tag-M1 +4μL Biotin-M2+ 6μL Protein L Donor 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 Streptavidin Acceptor Beads，Protect from Light/Green Light |
Add 6μL Streptavidin Acceptor 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 |
Readout |
Instrument Reading |
Instrument Reading |
[Performance Validation]
•Sample Preparation:
Use Universal Buffer 1 to pre-dilute Biotinylated Human IgG (Bio-hIgG) to 15μg/mL (100nM) as 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 L Donor Beads |
25 μg/mL |
Universal Buffer 1 |
Streptavidin Acceptor Beads |
25 μg/mL |
Universal Buffer 1 |
•37℃ Incubation Mode Detection Results:
Max Signal: 450369
Min Signal: 616
EC50= 0.082 nM
•Room Temperature Incubation Mode Detection Results:
Max Signal: 191706
Min Signal: 279
EC50= 0.059 nM
Guidelines
