ELISA Kit for Osteoprotegerin (OPG)

TNFRSF11B; TNFRSF11-B; OCIF; OPG; TR1; Tumor Necrosis Factor Receptor Superfamily Member 11b; Osteoclastogenesis Inhibitory Factor

Specificity

This assay has high sensitivity and excellent specificity for detection of Osteoprotegerin (OPG).
No significant cross-reactivity or interference between Osteoprotegerin (OPG) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Osteoprotegerin (OPG) and the recovery rates were calculated by comparing the measured value to the expected amount of Osteoprotegerin (OPG) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 99-105 102
EDTA plasma(n=5) 82-95 88
heparin plasma(n=5) 84-92 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Osteoprotegerin (OPG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Osteoprotegerin (OPG) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Osteoprotegerin (OPG) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 96-105% 90-98% 97-104% 94-102%
EDTA plasma(n=5) 88-101% 90-98% 83-99% 78-101%
heparin plasma(n=5) 80-101% 88-105% 98-105% 80-92%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Pharmacology Protective Effect of Amlodipine on Rat Bone Tissue after Orchidectomy Karger: 335491
Endocrine Pharmacology Protective effect of atorvastatin on bone tissue in orchidectomised male albino Wistar rats ScienceDirect: S0014299912000659
PLOS One Adipokines in Psoriatic Arthritis Patients: The Correlations with Osteoclast Precursors and Bone Erosions PubMed: PMC3483160
International Journal of Oral and Maxillofacial Surgery The effects of chronic zoledronate usage on the jaw and long bones evaluated using RANKL and osteoprotegerin levels in an animal model Pubmed: 23522850
American Journal of Orthodontics and Dentofacial Orthopedics OPG and RANKL levels around miniscrew implants during orthodontic tooth movement Pubmed: 23910201
AGE Estrogen deficiency does not decrease the in vitro osteogenic potential of rat adipose-derived mesenchymal stem cells Pubmed: 24687841
Food Science & Nutrition Effects and mechanisms of 8-prenylnaringenin on osteoblast MC3T3-E1 and osteoclast-like cells RAW264.7 Onlinelibrary: fsn3.109
Food Sci Nutr. Effects and mechanisms of 8‐prenylnaringenin on osteoblast MC3T3‐E1 and osteoclast‐like cells RAW264. 7 Pubmed:Pmc4221832
J Neurol Sci. The effect of topiramate and lamotrigine on rat bone mass, structure and metabolism. Pubmed:24629477
Journal of Investigative Surgery Effect of Nicotine on RANKL and OPG and Bone Mineral Density Pubmed:24830656
Int J Mol Med. Effects of icariin on the regulation of the OPG?RANKL?RANK system are mediated through the MAPK pathways in IL?1β?stimulated human SW1353 chondrosarcoma … Pubmed:25270538
Clinical and Experimental Pharmacology and Physiology Carvedilol promotes neurological function, reduces bone loss and attenuates cell damage after acute spinal cord injury in rats Pubmed:25424914
Journal of Colloid and Interface Science Strontium and zoledronate hydroxyapatites graded composite coatings for bone prostheses Pubmed:25706198
Int J Clin Exp Pathol A novel model of bisphosphonate-related osteonecrosis of the jaw in rats PubMed: 26191212
IJHS(international journal of health sciences) Relation of Osteoprotegerin, Visfatin and Ghrelin to Metabolic Syndrome in Type 2 Diabetic Patients PubMed: 26309431
Pharmacology Effect of Mirtazapine on Rat Bone Tissue after Orchidectomy PubMed: 25871861
Experimental And Therapeutic Medicine Antiosteoporotic effect of icariin in ovariectomized rats is mediated via the Wnt/β-catenin pathway Pubmed:27347050
Bone The effects of strength training and raloxifene on bone health in aging ovariectomized rats Pubmed:26812611
Scienze Biomediche Effetti del laser a bassa dose su cellule osteoblastiche INCERTIPARENTI_SERENA_TESI.pdf
journal of biomedical materials research part a RAW 264.7 co-cultured with ultra-high molecular weight polyethylene particles spontaneously differentiate into osteoclasts: an in vitro model of periprosthetic osteolysis. pubmed:27667508
Nutrition. Noni leaf and black tea enhance bone regeneration in estrogen-deficient rats. pubmed:27908549
PLoS One. Extracorporeal shock waves alone or combined with raloxifene promote bone formation and suppress resorption in ovariectomized rats. pubmed:28158228
Sci Rep. Skeletal Site-specific Effects of Zoledronate on in vivo Bone Remodeling and in vitro BMSCs Osteogenic Activity. pubmed:28139685
Journal of Biomaterials and Tissue Engineering Modification and Characterization of Polyethylene Glycolated Decellularized Porcine Aortic Heart Valve with Osteoprotegerin art00007
Japanese Society for Bone and Mineral Research Allogeneic yet major histocompatibility complex‑matched bone marrow transplantation in mice results in an impairment of osteoblasts and a significantly reduced trabecular bone article:10.1007
ONCOLOGY LETTERS Assessment of bone metabolism and biomechanical properties of the femur, following treatment with anastrozole and letrozole in an experimental model of menopause ol.2017.6596
Journal of Biomedical Materials Research RAW 264.7 co-cultured with ultra-high molecular weight polyethylene particles spontaneously differentiate into osteoclasts: an in vitro model of periprosthetic osteolysis 10.1002/jbm.a.35912
Oncotarget Mesenchymal stem cells inhibit RANK-RANKL interactions between osteoclasts and Th17 cells via osteoprotegerin activity. pubmed:29137353
Asian Journal of Biomedical and Pharmaceutical Sciences A Water Extract of Benefiting-Bone Capsule Improves Osteoporosis via Transcriptional and Translational Regulation of Key Factors in Notch and Wnt/β-catenin Signaling Pathways in Ovariectomized Rats ISSN 2249 – 622X
ACS Applied Materials and Interfaces Enhanced Osseointegration of Porous Titanium Modified with Zeolitic Imidazolate Framework-8. pubmed:28696091
Journal of Clinical Periodontology  Peroxisome proliferator‐activated receptor γ plays dual roles on experimental periodontitis in rats Pubmed:29574908
Journal of Functional Foods Effect of long-term administration of mangiferin from on bone metabolism in ovariectomized rats 10.1016:j.jff.2018.04.048
Macromolecular Bioscience Strontium‐Substituted Hydroxyapatite‐Gelatin Biomimetic Scaffolds Modulate Bone Cell Response Pubmed:29877029
Journal of Ethnopharmacology Effects of water extract from epimedium on neuropeptide signaling in an ovariectomized osteoporosis rat model Pubmed:29705515
International Journal of Molecular Medicine Bu‑Shen‑Ning‑Xin decoction suppresses osteoclastogenesis by modulating RANKL/OPG imbalance in the CD4+ T lymphocytes of ovariectomized mice Pubmed:29717766
Journal of Bone and Mineral Metabolism Allogeneic yet major histocompatibility complex-matched bone marrow transplantation in mice results in an impairment of osteoblasts and a significantly reduced … Pubmed:28766138
Journal of Bone and Mineral Metabolism  Therapeutic effect of icariin combined with stem cells on postmenopausal osteoporosis in rats Pubmed:28681147
Pharmacology Effects of Amlodipine on Bone Metabolism in Orchidectomised Spontaneously Hypertensive Rats Pubmed:29898457
Experimental and Therapeutic Medicine Liraglutide exerts a bone‑protective effect in ovariectomized rats with streptozotocin‑induced diabetes by inhibiting osteoclastogenesis Pubmed:29805533
Bone The sole and combined effect of simvastatin and platelet rich fibrin as a filling material in induced bone defect in tibia of albino rats Pubmed: 30208342
Bone Research NUMB maintains bone mass by promoting degradation of PTEN and GLI1 via ubiquitination in osteoblasts Pubmed: 30455992
Bone Development and characterization of supramolecular calcitonin assembly and assessment of its interactions with the bone remodelling process Pubmed: 30797058
RSC Advances Biofunctionalization of decellularized porcine aortic valve with OPG-loaded PCL nanoparticles for anti-calcification Doi: 10.1039/C9RA00408D
ANTICANCER RESEARCH Promoter Methylation Down-regulates Osteoprotegerin Expression in Ovarian Carcinoma Pubmed: 31092428
BIOSCIENCE REPORTS IL-38 restrains inflammatory response of collagen-induced arthritis in rats via SIRT1/HIF-1α signaling pathway Pubmed: 32347300
Oxid Med Cell Longev Long-Term Administration of Abacavir and Etravirine Impairs Semen Quality and Alters Redox System and Bone Metabolism in Growing Male Wistar Rats 34373766
Biomed Pharmacother Jiangu granule ameliorated OVX rats bone loss by modulating gut microbiota-SCFAs-Treg/Th17 axis Pubmed:35453007
Medicina (Kaunas) Plasma Concentrations of New Biochemical Markers of Atherosclerosis in Patients with Dyslipidemia—A Pilot Study Pubmed:35743980
Journal of Orthopaedic Translation Exercised accelerated the production of muscle-derived kynurenic acid in skeletal muscle and alleviated the postmenopausal osteoporosis through the Gpr35 …
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