OPG ELISA Kit (Human) (OKBB00235)

Catalog No: OKBB00235

Size:96 Tests

Human OPG ELISA Kit standard curve

Product Info

• Predicted Species Reactivity: Human
• Application: ELISA
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Duration: ~ 3 Hours
• ELISA Kit Principle: Aviva's human OPG ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for OPG has been precoated onto 96-well plates. Standards(NS0, E22-L401) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for OPG is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The absorbance of yellow is proportional to the human OPG amount of sample captured in plate.
• ELISA Kit Range: 93.7 pg/ml - 6,000 pg/ml
• ELISA Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	843	2186	3627	924	2570	4085
  St. Dev.	40.5	111.5	214	60.06	179.9	310.46
  %CV	4.80	5.10	5.90	6.5	7	7.6

• ELISA Kit Component:
  • Lyophilized recombinant human OPG standard: 10 ng/tube x 2
  • Anti-human OPG Antibody Well Plate: 96 Wells
  • Sample diluent buffer: 30 mL
  • Biotinylated anti-human OPG antibody: 130 uL, dilution 1:100
  • Antibody diluent buffer: 12 mL
  • Avidin-Biotin-Peroxidase Complex (ABC): 130 uL, dilution 1:100
  • ABC diluent buffer: 12 mL
  • TMB color developing agent: 10 mL
  • TMB stop solution: 10 mL
• Additional Information: Range: 93.8pg/ml-6000pg/ml
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
• Reconstitution and Storage: Store at 4C for 6 months, at -20C for 12 months. Avoid multiple freeze-thaw cycles (Shipped with wet ice.)
• Sample Type: cell culture supernates, serum, plasma(heparin, EDTA), saliva and urine
• Sensitivity: <5 pg/ml
• Reference: 1. Yasuda, H.; Shima, N.; Nakagawa, N.; Mochizuki, S. I.; Yano, K.; Fujise, N.; Sato, Y.; Goto, M.; Yamaguchi, K.;
  Kuriyama, M.; Kanno, T.; Murakami, A.; Tsuda, E.; Morinaga, T.; Higashio, K. Identity of osteoclastogenesis
  inhibitory factor (OCIF) and osteoprotegerin (OPG): a mechanism by which OPG/OCIF inhibits osteoclastogenesis in
  vitro. Endocrinology 139: 1329-1337, 1998.
  2. Ohmori, H.; Makita, Y.; Funamizu, M.; Hirooka, K.; Hosoi, T.; Orimo, H.; Suzuki, T.; Ikari, K.; Nakajima, T.; Inoue, I.;
  Hata, A. Linkage and association analyses of the osteoprotegerin gene locus with human osteoporosis. J. Hum.
  Genet. 47: 400-406, 2002.
  3. Kiechl, S.; Schett, G.; Wenning, G.; Redlich, K.; Oberhollenzer, M.; Mayr, A.; Santer, P.; Smolen, J.; Poewe, W.;
  Willeit, J. Osteoprotegerin is a risk factor for progressive atherosclerosis and cardiovascular disease. Circulation
  109: 2175-2180, 2004.
  4. Simonet, W. S.; Lacey, D. L.; Dunstan, C. R.; Kelley, M.; Chang, M.-S.; Luthy, R.; Nguyen, H. Q.; Wooden, S.;
  Bennett, L.; Boone, T.; Shimamoto, G.; DeRose, M.; and 19 others. Osteoprotegerin: a novel secreted protein
  involved in the regulation of bone density. Cell 89: 309-319, 1997.
  5. Croucher, P. I.; Shipman, C. M.; Lippitt, J.; Perry, M.; Asosingh, K.; Hijzen, A.; Brabbs, A. C.; van Beek, E. J. R.;
  Holen, I.; Skerry, T. M.; Dunstan, C. R.; Russell, G. R.; Van Camp, B.; Vanderkerken, K. Osteoprotegerin inhibits the development of osteolytic bone disease in multiple myeloma. Blood 98: 3534-3540, 2001.
  6. Brandstrom, H.; Stiger, F.; Lind, L.; Kahan, T.; Melhus, H.; Kindmark, A. A single nucleotide polymorphism in the promoter region of the human gene for osteoprotegerin is related to vascular morphology and function. Biochem. Biophys. Res. Commun. 293: 13-17, 2002.
• Specificity: Natural and recombinant human OPG
• Immunogen: Expression system for standard: NS0; Immunogen sequence: E22-L401
• Description: For quantitative detection of human OPG in cell culture supernatants, serum, plasma(heparin, EDTA), saliva and urine.

Target Info

• Gene Symbol: TNFRSF11B
• Gene Full Name: tumor necrosis factor receptor superfamily, member 11b
• Alias Symbols: MGC29565, MGC93568, OCIF, OPG, Osteoclastogenesis inhibitory factor, Osteoprotegerin, TNFRSF 11B, TNFRSF11B, TR 1, TR1, TR11B_HUMAN, Tumor necrosis factor receptor superfamily member 11B
• NCBI Gene Id: 4982
• Protein Name: Tumor necrosis factor receptor superfamily member 11B
• Description of Target: Osteoprotegerin (OPG) is identical to osteoclastogenesis inhibitory factor (OCIF), a soluble member of the tumor-necrosis factor receptor family that inhibits osteoclastogenesis.1 OPG is considered to play an important role in the regulation of bone resorption by modifying osteoclast differentiation.2 Osteoprotegerin is an independent risk factor for the progression of atherosclerosis and onset of cardiovascular disease.3 It can act as a soluble factor in the regulation of bone mass and imply a utility for OPG in the treatment of osteoporosis associated with increased osteoclast activity.4 OPG system may play a critical role in the development of osteolytic bone disease in multiple myeloma and that targeting this system may have therapeutic potential.5 OPG also plays a role in the vascular system.6 The standard product used in this kit is recombinant OPG, a 55KDa glycoprotein.
• Uniprot ID: O00300
• Protein Accession #: NP_002537.3
• Nucleotide Accession #: NM_002546.3