Nori Human Thioredoxin 1 (Trx1) ELISA Kit
$508.00 – $916.00
DataSheet Â
This ELISA kit is for quantification of thioredoxin 1 in human. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for Trx1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Trx1 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for Trx1 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of Trx1 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for Trx1: Thioredoxin 1, TXN
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Human Thioredoxin 1 (Trx1) ELISA Kit Summary
Alternative names for Trx1: Thioredoxin 1, TXN
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number | P10599 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 78 pg/mL |
| Detection Range | 0.391-25 ng/mL |
| Specificity | Human thioredoxin 1 (Trx1) |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:Â
Thioredoxin is a class of small redox proteins known to be present in all organisms. It acts as antioxidants by facilitating the reduction of other proteins by cysteine thiol-disulfide exchange and plays a role in many important biological processes, including redox signaling. Thioredoxins are encoded by TXN and TXN2 genes.[1] Loss-of-function mutation of either of the two thioredoxin genes is lethal at the four-cell stage of the developing embryo. Although not entirely understood, thioredoxin plays a central role and is increasingly linked to medicine through their response to reactive oxygen species (ROS). Trx1 has been shown to downregulate cardiac hypertrophy, the thickening of the walls of the lower heart chambers, by interactions with several different targets. Trx1 upregulates the transcriptional activity of nuclear respiratory factors 1 and 2 (NRF1 and NRF2) and stimulates the expression of peroxisome proliferator-activated receptor γ coactivator 1-α (PGC-1α).[2][3] Furthermore, Trx1 reduces two cysteine residues in histone deacetylase 4 (HDAC4), which allows HDAC4 to be imported from the cytosol, where the oxidized form resides,[4] into the nucleus.[5] Once in the nucleus, reduced HDAC4 downregulates the activity of transcription factors such as NFAT that mediate cardiac hypertrophy.[6] Trx 1 also controls microRNA levels in the heart and has been found to inhibit cardiac hypertrophy by upregulating miR-98/let-7.[7]
References
- Wollman EE, et al. (1988). The Journal of Biological Chemistry. 263 (30): 15506–12.
- Ago T, et al. (2006). Antioxidants & Redox Signaling. 8 (9–10): 1635–50.
- Yamamoto M, et al. (2003). The Journal of Clinical Investigation. 112 (9): 1395–406.
- Matsushima S, et al. (2013). Circulation Research. 112 (4): 651–63.
- Ago T, et al. (2008). Cell. 133 (6): 978–93.
- Nagarajan N, et al. (2016). Free Radical Biology & Medicine. 109: 125–131.
- Yang Y, et al. (2011). Circulation Research. 108 (3): 305–13.
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