Description
| Product Name: | Horse Transforming Growth Factor Beta-1 (TGFB1) ELISA Kit |
| Product Code: | EACO0003 |
| Product Size: | 96 Assays |
| Assay Type: | Sandwich ELISA |
| Detection Wavelength: | 450 nm |
| Range: | 62.5–4000 pg/mL |
| Sensitivity: | 15.6 pg/mL |
| Sample Types: | Serum, Plasma |
| Reactivity: | Equus caballus (Horse) |
| Intra-Assay CV: | <8% |
| Inter-Assay CV: | <10% |
| RUO: | Research-Use-Only |
This sandwich ELISA utilises a pre-coated antibody specific for TGFB1 on 96-well microplate. Standards and samples are added to wells, allowing any TGFB1 present to bind to immobilised antibody. After incubation, unbound substances are removed by washing. A biotin-conjugated detection antibody specific for TGFB1 is then added, which binds to captured target. Following wash to remove excess detection antibody, avidin-conjugated horseradish peroxidase (HRP) is introduced, forming biotin–avidin–HRP complex. After further 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 TGFB1 concentration, determined by reference to standard curve. This assay measures total TGFB1; latent TGFB1 must first be converted to immunoreactive form using acid-activation procedure.
| Standard Range: | 62.5–4000 pg/mL (2-fold dilution series, 7 standards + blank) |
| Sample Preparation: | Serum: Use SST, clot 2h RT or overnight 4°C, centrifuge 1000 × g 15 min; Plasma: EDTA or heparin, centrifuge 1000 × g 15 min at 2–8°C within 30 min |
| Sample Storage: | 2–8°C up to 5 days; -20°C ≤1 month; -80°C ≤2 months; avoid repeated freeze-thaw |
| Sample Activation (Required): | Add 20 µL 1 N HCl to 80 µL sample, mix well; incubate 10 min RT; neutralise with 16 µL 1.2 N NaOH / 0.5 M HEPES; mix well and assay immediately (final pH 7.2–7.6) |
| Standard/Sample Addition: | 100 µL per well |
| First Incubation: | 2 hours at 37°C, covered |
| First Wash: | Remove liquid without washing (no wash step) |
| Biotin-antibody (1×): | 100 µL per well, 1 hour at 37°C, covered |
| Wash Cycles: | 3 washes, 200 µL Wash Buffer per well, 2 min soak each |
| HRP-avidin (1×): | 100 µL per well, 1 hour at 37°C, covered |
| Wash Cycles: | 5 washes, 200 µL Wash Buffer per well |
| TMB Substrate: | 90 µL per well, 15–30 minutes at 37°C, protected from light |
| Stop Solution: | 50 µL per well, gently tap to mix; colour changes blue to yellow |
| Read: | 450 nm within 5 minutes after stop solution (correction at 540 nm or 570 nm if available) |
