Human Monoclonal Antibody Screening Set (GST-tag Antigen)
Human Monoclonal Antibody Screening Set (GST-tag Antigen)
Product Details
Product Details
Product Specification
| Stability & Storage | Store at 2～8℃ 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 to generate 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 high speed and sensitivity. It is capable of detecting weak interactions.
Components
Name |
Main Component |
3000T |
Reagent A |
Anti-GST-tag antibody-crosslinked acceptor Beads |
10mL |
Reagent B |
Biotin-labeled anti-human IgG antibody |
10mL |
Reagent D |
Streptavidin Donor Beads |
10mL |
Protocol
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【Required Reagents】
Name |
Catalog Number |
Universal Buffer 4 |
UA086115 |
【Detection Procedure (For Reference)】
Detection Step |
Procedure |
Step 1: |
Add GST-tag antigen to Reagent A at a concentration of 2μg/mL |
Step 2: |
Mix Reagent A, Reagent B, and Reagent D uniformly at a volume ratio of 1:1:1,protect from light/green light |
Step 3: |
Add 10μL of test sample + 10μL of mixed reagent per test,protect from light/green light |
Incubation |
37°C with shaking for 60 minutes,protect from light/green light |
Reading |
Instrument reading |
【Sample and Reagent Preparation】
•Pre-dilute H-1b antibody to 1000ng/mL (6.67nM) using Universal Buffer 4 as C8, then perform gradient dilution as follows:
ID |
Concentration (ng/mL) |
Universal Buffer 4 Volume (μL) |
High Concentration Addition Volume (μL) |
C8 |
1000 |
/ |
/ |
C7 |
200 |
160 |
40μL C8 |
C6 |
40 |
160 |
40μL C7 |
C5 |
8 |
160 |
40μL C6 |
C4 |
1.6 |
160 |
40μL C5 |
C3 |
0.32 |
160 |
40μL C4 |
C2 |
0.064 |
160 |
40μL C3 |
C1 |
0 |
Guidelines
1. This experiment is light-sensitive. Ensure all operations are performed under light-protected conditions. It is recommended to complete preparation, sample loading, and incubation steps under green light (illuminance below 100 LUX).
2. This product is compatible with multifunctional microplate readers equipped with Alpha detection modules.
3. To ensure comparability of experimental data across different batches, strictly control incubation temperature and duration.
4. Avoid generating bubbles during sample loading.
