Description

Nori Hamster IL-6 ELISA Kit Summary

Alternative names for IL-6: Interleukin 6, IL6

Alternative names for hamster: Golden hamster, Syrian hamster, Chinese hamster

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