Description
| Product Name: | Horse TGF-b1 (Transforming Growth Factor Beta 1) ELISA Kit (Activation Required) |
| Product Code: | AEKE13070 |
| Product Size: | 48T/96T |
| Assay Type: | Sandwich ELISA |
| Detection Wavelength: | 450 nm |
| Range: | 0.16-10 ng/mL |
| Sensitivity: | 0.11 ng/mL |
| Sample Types: | Cell Culture Supernatant, Cell Lysate, Other Biological Fluids, Plasma, Serum, Tissue Homogenate |
| Intra-Assay CV: | <8% |
| Inter-Assay CV: | <10% |
| RUO: | Research-Use-Only |
This sandwich ELISA employs a pre-coated antibody specific for target protein on 96-well microplate. Samples and standards are added to wells, allowing target to bind to immobilized antibody. After incubation, unbound substances are removed by washing. A biotinylated detection antibody is then added, which binds specifically to captured target. Following wash to remove excess detection antibody, HRP-conjugated Streptavidin is introduced, forming biotin-streptavidin-HRP complex. After third washing step, TMB substrate is added to initiate colorimetric reaction catalysed by HRP, producing blue product that turns yellow upon addition of acidic Stop Solution. OD450 is directly proportional to target concentration, determined by referencing standard curve.
| Standard Range: | 0.16-10 ng/mL (7-point serial dilution) |
| Sample Preparation: | Serum (2h coagulation, centrifuge 1000 × g 20 min); Plasma (EDTA/citrate/heparin, centrifuge 1000 × g 15 min); Tissue (homogenize 1:9 w/v PBS + protease inhibitor, centrifuge 5000 × g 5 min); Cell lysate (10,000 rpm 10 min) |
| Sample Activation (Required): | Add 20 µL sample to 140 µL Diluent; add 20 µL Activator A, incubate 10 min RT; add 20 µL Activator B, mix well; test immediately (10-fold dilution) |
| Standard/Sample Addition: | 100 µL per well to bottom |
| First Incubation: | 37°C, 80 minutes, sealed |
| Wash Cycles: | 3 washes, 200 µL 1X Wash Buffer per well, 1-2 min soak each |
| Biotinylated Antibody: | 100 µL per well, 37°C 50 minutes |
| Wash Cycles: | 5 washes, 200 µL 1X Wash Buffer per well |
| Streptavidin-HRP: | 100 µL per well, 37°C 50 minutes |
| Wash Cycles: | 5 washes, 200 µL 1X Wash Buffer per well |
| TMB Substrate: | 90 µL per well, 37°C 20 minutes in dark (do not exceed 30 minutes) |
| Stop Solution: | 50 µL per well; colour changes blue to yellow immediately |
| Read: | 450 nm immediately after stop solution |
