Description

Nori Mouse IL-6 ELISA Kit Summary

Alternative names for IL-6: IL6, interleukin 6

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: 
IL-6 is an interleukin that acts as both a pro-inflammatory and anti-inflammatory cytokine and is produced by T cells, macrophages, fibroblasts, osteoblasts, endothelial and other cells (1,2,3). IL-6 induces proliferation and differentiation and acts on B cells, T cells, thymocytes, and others. IL-6 is one of the most important mediators of fever and of the acute phase response. In the muscle and fatty tissue, IL-6 stimulates energy mobilization that leads to increased body temperature. IL-6 can be secreted by macrophages in response to specific microbial molecules, referred to as pathogen associated molecular patterns (PAMPS). IL-6 in concert with TGFβ is important for developing Th17 responses. IL-6 binds to IL-6Rα that through association induces gp130 homodimerization (1). gp130 homodimerization triggers the Jak/STAT cascade and the SHP2/Erk Map kinase cascade (1,4,5). IL-6 also forms a complex with an IL-6Rα splice variant that is non-membrane associated (4). The IL-6/soluble IL-6Rα complex can then activate the gp130 signaling pathway on cells that express gp130 but not IL6Rα (4). IL-6 is relevant to many disease processes such as diabetes (6), atherosclerosis (7), depression (8), Alzheimer’s Disease (9), systemic lupus erythematosus (10), prostate cancer (11), breast cancer (12), and rheumatoid arthritis (13).

References
1. Heinrich, P.C. et al. (1998) Biochem J 334 ( Pt 2), 297-314.
2. Heinrich, P.C. et al. (1998) Z Ernahrungswiss 37 Suppl 1, 43-9.
3. Febbraio MA and Pedersen BK (2005). Exerc Sport Sci Rev 33 (3): 114–9.
4. Jones, S.A. (2005) J Immunol 175, 3463-8.
5. Jenkins, B.J. et al. (2004) Mol Cell Biol 24, 1453-63.
6. Kristiansen OP and Mandrup-Poulsen T (2005). Diabetes 54 Suppl 2: S114–24.
7. Dubiński A and Zdrojewicz Z (2007). Pol. Merkur. Lekarski 22 (130): 291–4.
8. Dowlati Y, et al (2010). Biological Psychiatry 67 (5): 446–457.
9. Swardfager W, et al (2010). Biological Psychiatry 68 (10): 930–941.
10. Tackey E, et al (2004). Lupus 13 (5): 339–43.
11. Smith PC, et al (2001). Cytokine Growth Factor Rev. 12 (1): 33–40.
12. Hong, D.S. et al. (2007) Cancer 110, 1911-28.
13. Nishimoto N (2006). Curr Opin Rheumatol 18 (3): 277–81

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).