Nori Rat LIGHT/TNFSF14 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of LIGHT/TNFSF14 in rat. 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 LIGHT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any LIGHT present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for LIGHT 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 LIGHT bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for LIGHT/TNFSF14: Tumor necrosis factor ligand superfamily member 14, HVEML
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Rat LIGHT/TNFSF14 ELISA Kit Summary
Alternative names for LIGHT/TNFSF14: Tumor necrosis factor ligand superfamily member 14, HVEML
| 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 | A0ACM8RBP8 |
| 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 | 18 pg/mL |
| Detection Range | 93.75-6000 pg/mL |
| Specificity | Rat LIGHT |
| 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
LIGHT, also known as tumor necrosis factor superfamily member 14 (TNFSF14), is a secreted protein of the TNF superfamily.[1] It is recognized by herpesvirus entry mediator (HVEM), as well as decoy receptor 3. LIGHT stands for “homologous to lymphotoxin, exhibits inducible expression and competes with HSV glycoprotein D for binding to herpesvirus entry mediator, a receptor expressed on T lymphocytes”. In the cluster of differentiation terminology it is classified as CD258. LIGHT is a ligand for TNFRSF14, which is also known as a herpesvirus entry mediator ligand (HVEML). LIGHT may function as a costimulatory factor for the activation of lymphoid cells and as a deterrent to infection by herpesvirus. LIGHT has been shown to stimulate the proliferation of T cells, and trigger apoptosis of various tumor cells. LIGHT is also reported to prevent tumor necrosis factor alpha mediated apoptosis in primary hepatocyte. Two alternatively spliced transcript variant encoding distinct isoforms have been reported. LIGHT has been shown to interact with TNFRSF14,[2][3] TNFRSF6B,[4] BIRC2,[5] TRAF2[5] and TRAF3.[5] Similar to how CD4 is the primary mediating receptor in HIV infection, the HSV glycoprotein (gD) binds to the HVEM receptor which is demanded by TNFSF14/LIGHT lowering the ability for LIGHT to activate the NFκB pathway. NFκB is a survival factor helping to inhibit apoptosis which triggers a pathway inhibiting caspase 8. When gD from HSV binds to HVEM, LIGHT is non-competitively inhibited from binding, encouraging apoptosis in the infected cell.
References
- Mauri DN, et al. (1998). Immunity. 8 (1): 21–30.
- Zhang J, et al. (2001). The Journal of Clinical Investigation. 107 (11): 1459–68.
- Yu KY, et al. (1999). The Journal of Biological Chemistry. 274 (20): 13733–6.
- Hsu TL, et al. (2002). Journal of Immunology. 168 (10): 4846–53.
- Kuai J, et al. (2003). The Journal of Biological Chemistry. 278 (16): 14363–9.
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