Human Osteoprotegerin (OPG) ELISA

RayBio Human Osteoprotegerin ELISA Kit for serum, plasma, cell culture supernatants, and urine

96-well Microplate

Quantitative

Cell Culture Supernatants, Plasma, Serum, Urine

1 pg/ml - 900 pg/ml

This ELISA kit shows no cross-reactivity with any of the cytokines tested (e.g., human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-11, IL-12 p70, IL- 12 p40, IL-13, IL-15, IL-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIP-1alpha, MIP-1 beta, MIP-1gamma, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF).

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 ul of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4C.
4. Add 100 ul of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 ul of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 ul of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 ul of Stop Solution to each well.
11. Read at 450 nm immediately.

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2. Maksymowych WP, Landewe R, Conner-Spady B, Dougados, et al. Serum Matrix Metalloproteinase 3 Is an Independent Predictor of Structural Damage Progression in Patients with Ankylosing Spondylitis. Arthritis Rheum. 2007, 56(6):1846–1853.
3. Jose´ I Botella-Carretero, Francisco Alvarez-Blasco, Jose´ Luis San Milla´n and He´ctor F Escobar-Morreale. Thyroid hormone deficiency and postmenopausal status independently increase serum osteoprotegerin concentrations in women. Eur J Endocrinol. 2007, 156:539–545
4. Nishigaki M, Yamamoto T, Ichioka H, et al. beta-cryptoxanthin regulates bone resorption related-cytokine production in human periodontal ligament cells. Arch Oral Biol. 2013 Jul, 58(7):880–886.
5. Akelma AZ, Cizmeci MN, Kanburoglu MK, Bozhkaya D, Catal F, et al. Elevated level of serum osteopontin in school-age children with asthma. Allergol Immunop (Madr). 2013, Published online ahead of print, April 8, 2013. doi:10.1016/j.aller.2013.01.009
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7. Hartwich JE, Orr WS, Ng CY, et al. Rapamycin increases neuroblastoma xenograft and host stromal derived osteoprotegerin inhibiting osteolytic bone disease in a bone metastasis model. J Pediatr Surg. 2013 Jan, 48(1):47–55.
8. Lucas R., Ramos E., Prata M., Rodrigues A., Costa L., et al. Changes in serum RANKL and OPG with sexual development and their associations with bone turnover and bone mineral density in a cohort of girls. Clinical Biochemistry Available online 22 April 2014 [Epub ahead of Print]
9. Palumbo S, Li W-J. Osteoprotegerin Enhances Osteogenesis of Human Mesenchymal Stem Cells. Tissue Eng Part A. Volume 19, Numbers 19 and 20, 2013, DOI: 10.1089/ten.tea.2012.0550
10. Holt V, Caplan AI, Haynesworth SE (2014) Identification of a Subpopulation of Marrow MSC-Derived Medullary Adipocytes That Express Osteoclast- Regulating Molecules: Marrow Adipocytes Express Osteoclast Mediators. PLoS ONE 9(10): e108920. doi:10.1371/journal.pone.0108920