Multiplex Assay Kit for Osteoprotegerin (OPG) ,etc. by FLIA (Flow Luminescence Immunoassay) Rattus norvegicus (Rat) Multiplex ELISA

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

(Note: Up to 8-plex in one testing reaction)

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  • Multiplex Assay Kit for Osteoprotegerin (OPG) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Osteoprotegerin (OPG) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Osteoprotegerin (OPG) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA108Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 90-97 94
EDTA plasma(n=5) 79-102 85
heparin plasma(n=5) 80-88 83
sodium citrate plasma(n=5) 88-96 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Osteoprotegerin (OPG) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Osteoprotegerin (OPG) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 95-103% 95-102% 85-101% 96-105%
EDTA plasma(n=5) 80-89% 97-105% 78-105% 78-96%
heparin plasma(n=5) 85-105% 80-90% 99-105% 89-104%
sodium citrate plasma(n=5) 91-99% 84-103% 79-104% 89-97%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:OPG) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

Multiplex Assay Kit for Osteoprotegerin (OPG) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Protective Effect of Amlodipine on Rat Bone Tissue after OrchidectomyKarger: 335491
  • Protective effect of atorvastatin on bone tissue in orchidectomised male albino Wistar ratsScienceDirect: S0014299912000659
  • Adipokines in Psoriatic Arthritis Patients: The Correlations with Osteoclast Precursors and Bone ErosionsPubMed: PMC3483160
  • The effects of chronic zoledronate usage on the jaw and long bones evaluated using RANKL and osteoprotegerin levels in an animal modelPubmed: 23522850
  • OPG and RANKL levels around miniscrew implants during orthodontic tooth movementPubmed: 23910201
  • Estrogen deficiency does not decrease the in vitro osteogenic potential of rat adipose-derived mesenchymal stem cellsPubmed: 24687841
  • Effects and mechanisms of 8-prenylnaringenin on osteoblast MC3T3-E1 and osteoclast-like cells RAW264.7Onlinelibrary: fsn3.109
  • Effects and mechanisms of 8‐prenylnaringenin on osteoblast MC3T3‐E1 and osteoclast‐like cells RAW264. 7Pubmed:Pmc4221832
  • The effect of topiramate and lamotrigine on rat bone mass, structure and metabolism.Pubmed:24629477
  • Effect of Nicotine on RANKL and OPG and Bone Mineral DensityPubmed:24830656
  • 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
  • Carvedilol promotes neurological function, reduces bone loss and attenuates cell damage after acute spinal cord injury in ratsPubmed:25424914
  • Strontium and zoledronate hydroxyapatites graded composite coatings for bone prosthesesPubmed:25706198
  • A novel model of bisphosphonate-related osteonecrosis of the jaw in ratsPubMed: 26191212
  • Relation of Osteoprotegerin, Visfatin and Ghrelin to Metabolic Syndrome in Type 2 Diabetic PatientsPubMed: 26309431
  • Effect of Mirtazapine on Rat Bone Tissue after OrchidectomyPubMed: 25871861
  • Antiosteoporotic effect of icariin in ovariectomized rats is mediated via the Wnt/β-catenin pathwayPubmed:27347050
  • The effects of strength training and raloxifene on bone health in aging ovariectomized ratsPubmed:26812611
  • Effetti del laser a bassa dose su cellule osteoblasticheINCERTIPARENTI_SERENA_TESI.pdf
  • 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
  • Noni leaf and black tea enhance bone regeneration in estrogen-deficient rats.pubmed:27908549
  • Extracorporeal shock waves alone or combined with raloxifene promote bone formation and suppress resorption in ovariectomized rats.pubmed:28158228
  • Skeletal Site-specific Effects of Zoledronate on in vivo Bone Remodeling and in vitro BMSCs Osteogenic Activity.pubmed:28139685
  • Modification and Characterization of Polyethylene Glycolated Decellularized Porcine Aortic Heart Valve with Osteoprotegerinart00007
  • Allogeneic yet major histocompatibility complex‑matched bone marrow transplantation in mice results in an impairment of osteoblasts and a significantly reduced trabecular bonearticle:10.1007
  • Assessment of bone metabolism and biomechanical properties of the femur, following treatment with anastrozole and letrozole in an experimental model of menopauseol.2017.6596
  • RAW 264.7 co-cultured with ultra-high molecular weight polyethylene particles spontaneously differentiate into osteoclasts: an in vitro model of periprosthetic osteolysis10.1002/jbm.a.35912
  • Mesenchymal stem cells inhibit RANK-RANKL interactions between osteoclasts and Th17 cells via osteoprotegerin activity.pubmed:29137353
  • 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 RatsISSN 2249 – 622X
  • Enhanced Osseointegration of Porous Titanium Modified with Zeolitic Imidazolate Framework-8.pubmed:28696091
  • Peroxisome proliferator‐activated receptor γ plays dual roles on experimental periodontitis in ratsPubmed:29574908
  • Effect of long-term administration of mangiferin from on bone metabolism in ovariectomized rats10.1016:j.jff.2018.04.048
  • Strontium‐Substituted Hydroxyapatite‐Gelatin Biomimetic Scaffolds Modulate Bone Cell ResponsePubmed:29877029
  • Effects of water extract from epimedium on neuropeptide signaling in an ovariectomized osteoporosis rat modelPubmed:29705515
  • Bu‑Shen‑Ning‑Xin decoction suppresses osteoclastogenesis by modulating RANKL/OPG imbalance in the CD4+ T lymphocytes of ovariectomized micePubmed:29717766
  • Allogeneic yet major histocompatibility complex-matched bone marrow transplantation in mice results in an impairment of osteoblasts and a significantly reduced …Pubmed:28766138
  • Therapeutic effect of icariin combined with stem cells on postmenopausal osteoporosis in ratsPubmed:28681147
  • Effects of Amlodipine on Bone Metabolism in Orchidectomised Spontaneously Hypertensive RatsPubmed:29898457
  • Liraglutide exerts a bone‑protective effect in ovariectomized rats with streptozotocin‑induced diabetes by inhibiting osteoclastogenesisPubmed:29805533
  • The sole and combined effect of simvastatin and platelet rich fibrin as a filling material in induced bone defect in tibia of albino ratsPubmed: 30208342
  • NUMB maintains bone mass by promoting degradation of PTEN and GLI1 via ubiquitination in osteoblastsPubmed: 30455992
  • Development and characterization of supramolecular calcitonin assembly and assessment of its interactions with the bone remodelling processPubmed: 30797058
  • Biofunctionalization of decellularized porcine aortic valve with OPG-loaded PCL nanoparticles for anti-calcificationDoi: 10.1039/C9RA00408D
  • Promoter Methylation Down-regulates Osteoprotegerin Expression in Ovarian CarcinomaPubmed: 31092428
  • IL-38 restrains inflammatory response of collagen-induced arthritis in rats via SIRT1/HIF-1α signaling pathwayPubmed: 32347300
  • Long-Term Administration of Abacavir and Etravirine Impairs Semen Quality and Alters Redox System and Bone Metabolism in Growing Male Wistar Rats34373766
  • Jiangu granule ameliorated OVX rats bone loss by modulating gut microbiota-SCFAs-Treg/Th17 axisPubmed:35453007
  • Plasma Concentrations of New Biochemical Markers of Atherosclerosis in Patients with Dyslipidemia—A Pilot StudyPubmed:35743980
  • Exercised accelerated the production of muscle-derived kynurenic acid in skeletal muscle and alleviated the postmenopausal osteoporosis through the Gpr35 …

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