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) 85-101 96
EDTA plasma(n=5) 82-101 89
heparin plasma(n=5) 96-104 101

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) 96-104% 80-96% 85-99% 78-104%
EDTA plasma(n=5) 97-104% 78-93% 97-105% 90-101%
heparin plasma(n=5) 89-103% 96-104% 80-92% 83-90%

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
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