Nori Rat RANKL ELISA Kit
$461.00 – $832.00
DataSheet CoA SDS
This ELISA kit is for quantification of RANKL 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 RANKL has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any RANKL present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for RANKL 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 RANKL bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for RANKL: Receptor activator of nuclear factor kappa-B ligand (RANKL), TNFSF11
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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- Product Citation (25)
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Description
Nori Rat RANKL ELISA Kit Summary
Alternative names for RANKL: Receptor activator of nuclear factor kappa-B ligand (RANKL), TNFSF11
| 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 | Q9ESE2 |
| 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 | 12 pg/mL |
| Detection Range | 62.5-4000 pg/mL |
| Specificity | Natural and recombinant rat RANKL |
| 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:
Receptor activator of nuclear factor kappa-B ligand (RANKL) is a protein that is encoded by the TNFSF11 gene.[1] RANKL is known as a type II membrane protein and is a member of the tumor necrosis factor (TNF) superfamily.[2] RANKL has been identified to affect the immune system and control bone regeneration and remodeling. RANKL may be used as an apoptosis regulator gene, a binding partner of OPG, a ligand for the receptor RANK and control proliferation by modifying protein levels of Id4, Id2 and cyclin D1.[3] RANKL is expressed in several tissues and organs and high protein expression of RANKL are commonly detected in the lung, thymus and lymph nodes.[3] The level of RANKL expression does not linearly correlate to the effect of this ligand., While bone marrow expresses low levels of RANKL, it plays a critical role for adequate bone metabolism, this surface-bound molecule (also known as CD254) found on osteoblasts serves to activate osteoclasts, which are critically involved in bone resorption. Osteoclastic activity is triggered via the osteoblasts’ surface-bound RANKL activating the osteoclasts’ surface-bound receptor activator of nuclear factor kappa-B (RANK). RANKL also has a function in the immune system, where it is expressed by T helper cells and is thought to be involved in dendritic cell maturation. This protein was shown to be a dendritic cell survival factor and is involved in the regulation of T cell-dependent immune response. T cell activation was reported to induce expression of this gene and lead to an increase of osteoclastogenesis and bone loss. This protein was shown to activate antiapoptotic kinase AKT/PKB through a signaling complex involving SRC kinase and tumor necrosis factor receptor-associated factor 6 (TRAF6), which indicated this protein may have a role in the regulation of cell apoptosis.
References
- Anderson DM, et al. (1997). Nature 390 (6656): 175–9.
- Hanada R, et al. (2011). J. Mol. Med. 89 (7): 647–56.
- Wada T, et al. (2006). Trends Mol Med 12 (1): 17–25.
Product Citation
40. Zucca E et al. (2016) Evaluation of amniotic mesenchymal cell derivatives on cytokine production in equine
alveolar macrophages: an in vitro approach to lung inflammation. Stem Cell Research and Therapy 7:137. Article
Product used and cited: Nori Equine TNFα/IL-6/TGFβ1 ELISA kits. Impact factor: 4.731.
91. Witonsky S et al. (2019) Can levamisole upregulate the equine cell-mediated macrophage (M1) dendritic
cell (DC1) T-helper 1 (CD4 Th1) T-cytotoxic (CD8) immune response in vitro? J Vet Internal Med 33(2):889-96.
Impact factor: 2.286.
Products used and cited: Nori Equine IL-6 ELISA Kit.
37. Siemieniuch MJ et al. (2016) Type of inflammation differentially affects expression of interleukin 1β and 6, tumor
necrosis factor-α and toll-like receptors in subclinical endometritis in mares. PLOS ONE 11(5):e0154934. Doi:10.137
/journal.pone.0154934. Impact factor: 2.806.
Product used and cited: Nori Equine IL-6 ELISA kit.
48. Water EVD et al. (2016) The preventive effects of two nutraceuticals on experimentally induced acute synovitis. Equine Veterinary Journal DOI:10.1111/evj.12629. Impact factor: 2.40.
Product used and cited: Nori Equine IL-6 ELISA Kit GR106001, GR106088.
81. Abass M et al. (2018) Local mepivacaine before castration of horses under medetomidine
isoflurane balanced anesthesia is effective to reduce perioperative nociception and cytokine release.
Equine Veterinary Journal, 50:733-738. Impact factor: 2.022
Produce used and cited: Nori Equine IL-6 and TNF-alpha ELISA Kit.
96. Broeckx SY et al. (2019) The use of equine chondrogenic-induced mesenchymal stem cells as a treatment for osteoarthritis:
A randomized, double-blinded, placebo-controlled proof-of-concept study. Equine Vet J 51(6): 787-94. Impact factor: 2.022.
Products used and cited: Nori Equine IL-6 and TGFβ3 ELISA Kits.
110. Hussein H et al. (2016) Cathepsin K inhibition renders equine bone marrow nucleated cells hypo-responsive
to LPS and unmethylated CpG stimulation in vitro. Comparative Immunology, Microbiology and Infectious Diseases
45: 40-7. Impact factor: 2.05.
Products used and cited: Nori Equine IL-1β, IL-6 and TNFα ELISA Kits.
126. Broeckx SY et al. (2019) Evaluation of an osteochondral fragment-groove procedure for induction of
metacarpophalangeal joint osteoarthritis in horses. Am J Vet Res 80(3):246-58. Impact factor: 1.07.
Products used and cited: Nori Equine IL-6 and TGFb3 ELISA Kits (GR106001, GR106134).
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