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Scientific Background
This ELISA kit applies to the in vitro quantitative determination of TGF-β1 concentrations in Serum, plasma and other biological fluids.
Assay Principle
This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Universal TGF-β1. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Universal TGF-β1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Universal TGF-β1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Universal TGF-β1. You can calculate the concentration of Universal TGF-β1 in the samples by comparing the OD of the samples to the standard curve.
Performance Specifications
| Sensitivity | 0.1 ng/mL |
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| Detection Range | 0.16-10 ng/mL |
| Total Assay Time | 3 h 30 min |
| Compatible Sample Types | Serum, plasma and other biological fluids |
| Species Reactivity | Universal (multi-species validated) |
| Detection Method | Sandwich |
| Precision (CV) | Both intra-CV and inter-CV are < 10%. |
| Recovery Rate | 80%-120% |
| Storage | 2-8℃,12 months |
✓ Research-Grade Validation
Specificity
This kit recognizes Universal TGF-β1 in samples.No significant cross-reactivity or interference between Universal TGF-β1 and analogues was observed
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with Serum, plasma and other biological fluids. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 0.1 ng/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3 h 30 min, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant TGF-β1 standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
Since the antibodies in the kit recognize activated TGF-β, and TGF-β is a disulfide bond linked by two identical or similar 12.5kDa subunits of molecular weight. The study of human TGF-β cDNA sequence showed that the 112 amino acid residues of the monomer TGF-β were cleaved from the carboxyl terminal by a precursor molecule (per-pro-TGF-β) containing 400 amino acid residues. The N-terminal of per-pro-TGF-β contains a signal peptide, which is cleaved before secretion to become an inactive polypeptide chain precursor (pro-TGF-β). The n-terminal part of the amino acid residue is removed by changing the ionic strength, acidification or protease hydrolysis, and the remaining carboxyl terminal part forms an active TGF-β. A variety of cells in the body can secrete TGF-β in an inactive state. In vitro, the inactive TGF-β, also known as latency associated peptide (LAP), can be activated by acidification. In vivo, the acidic environment can be present near fractures and healing wounds, and the cleavage of the protein itself can cause the TGF-β complex to become activated TGF-β. In general, tissues with active cell differentiation often contain high levels of TGF-β, such as osteoblasts, kidney, bone marrow, and fetal liver hematopoietic cells. Therefore, no additional activation processing is required for tissue sample testing.
As long as the samples and reagents used during the activation step are consistent with the proportions in the manual, it should fine.
Why is it necessary to add a protease inhibitor in tissue sample preparation during an Elisa experiment? Will it affect the detection significantly if there is no protease inhibitor?
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