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ELISA Kit for Osteopontin (OPN) Mus musculus (Mouse) Sandwich ELISA

SPP1; BNSP; BSPI; ETA1; Secreted Phosphoprotein 1; Bone Sialoprotein I; Early T-Lymphocyte Activation 1; Nephropontin; Urinary stone protein; Uropontin

  • Product No.SEA899Mu
  • Organism SpeciesMus musculus (Mouse) Same name, Different species.
  • Sample Typeserum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
  • Test MethodDouble-antibody Sandwich
  • Assay Length3h
  • Detection Range39-2,500pg/mL
  • SensitivityThe minimum detectable dose of this kit is typically less than 15pg/mL.
  • Specificity This assay has high sensitivity and excellent specificity for detection of Osteopontin (OPN).No significant cross-reactivity or interference between Osteopontin (OPN) and analogues was observed.
  • ApplicationsEnzyme-linked immunosorbent assay for Antigen Detection.
  • DownloadInstruction Manual
  • UOM 48T96T 96T*5 96T*10 96T*100 Ensure in stock; if not, free redo!
  • FOB US$ 349.00 US$ 498.00 US$ 2,241.00 US$ 4,233.00 US$ 34,860.00
  • Quantity
  • Add to Cart Distributors
  • ELISA Kit for Osteopontin (OPN) Packages (Simulation)
  • ELISA Kit for Osteopontin (OPN) Packages (Simulation)
  • ELISA Kit for Osteopontin (OPN) Results demonstration
  • SEA899Mu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 89-101 96
EDTA plasma(n=5) 80-99 85
heparin plasma(n=5) 99-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Osteopontin (OPN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Osteopontin (OPN) 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 Osteopontin (OPN) 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) 87-96% 89-96% 79-91% 96-103%
EDTA plasma(n=5) 88-95% 87-97% 92-101% 80-94%
heparin plasma(n=5) 92-105% 80-104% 90-104% 80-94%

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.

ELISA Kit for Osteopontin (OPN)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Osteopontin (OPN). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Osteopontin (OPN). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Osteopontin (OPN), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Osteopontin (OPN) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reference

  • Journal of Biomedical Materials Research Part AProliferation and osteogenesis of immortalized bone marrow-derived mesenchymal stem cells in porous polylactic glycolic acid scaffolds under perfusion culturePubMed: 19280635
  • Osteoarthritis and CartilageElevated osteopontin level of synovial fluid and articular cartilage is associated with disease severity in knee osteoarthritis patientsPubMed: 19747583
  • The Journal of RheumatologyThrombin-cleaved osteopontin levels in synovial fluid correlate with disease severity of knee osteoarthritis.Jrheum: source
  • International Journal of PharmaceuticsSuppression of tumor growth in xenograft model mice by small interfering RNA targeting osteopontin delivery using biocompatible poly(amino ester)ScienceDirect: S0378517312003742
  • European Cells and matreialsThe use of non-viral gene vectors for bioactive poly-(D, L-lactide) implant surfaces in bone tissue engineeringEcmjournal: Source
  • Journal of Equine Veterinary ScienceInfluence of Seminal Plasma Antioxidants and Osteopontin on Fertility of the Arabian HorseScienceDirect: Source
  • Basic &amp; Clinical Pharmacology &amp; ToxicologyTreatment with Carnitine Enhances Bone Fracture Healing under Osteoporotic and/or Inflammatory ConditionsWiley:Source
  • Journal of Cellular BiochemistryTension Force-Induced ATP Promotes Osteogenesis Through P2X7 Receptor in OsteoblastsPubmed:24905552
  • Naunyn-Schmiedeberg&#039;s Archives of PharmacologyTreatment with α-lipoic acid enhances the bone healing after femoral fracture model of ratsPubmed:25038619
  • J Endocrinol Invest.?The interplay among iron metabolism, endothelium and inflammatory cascade in dysmetabolic disordersPubmed:25245337
  • FEBS LettersLow-intensity pulsed ultrasound-induced ATP increases bone formation via the P2X7 receptor in osteoblast-like MC3T3-E1 cellsPubmed:25542352
  • GeneEffects of magnesium degradation products on mesenchymal stem cell fate and osteoblastogenesisPubMed: 26283150
  • Medicine (Baltimore)An Attempt to Evaluate Selected Aspects of “Bone–Fat Axis” Function in Healthy Individuals and Patients With Pancreatic CancerPubMed: 26266370
  • Journal of Biomaterials and Tissue EngineeringOsteogenic Differentiation of Canine Adipose Stem Cells Cultured in Alginate-Fibrin-Based HydrogelContent: Asp
  • PrimasariOsteopontin level correlates negatively with tumor shrinkage in neoadjuvant chemoradiation of locally advanced rectal cancerIndex.Php: Amor
  • Int J Biol SciDefective Initiation of Liver Regeneration in Osteopontin-Deficient Mice after Partial Hepatectomy due to Insufficient Activation of IL-6/Stat3 PathwayPubMed: 26327817
  • Journal of Kunming Medical University / Kunming Yike Daxue XuebaoPerioperative Clinical Investigation of Serum VEGF, CA19-9 and OPN Levels in Patients with Colorectal Carcinoma.Ebscohost
  • Am J Pathol. Serum Osteopontin as a Novel Biomarker for Muscle Regeneration in Duchenne Muscular Dystrophy.Pubmed:26963343
  • circulation journalStenotic Bicuspid and Tricuspid Aortic Valves - Micro-Computed Tomography and Biological Indices of Calcification.pubmed:28344201
  • Journal of Cardiovascular ElectrophysiologyEffect of Cardiac Resynchronization Therapy on Myocardial Fibrosis and Relevant Cytokines in a Canine Model With Experimental Heart Failure.pubmed:28127817
  • American Journal of Translational ResearchOsteopontin stimulates matrix metalloproteinase expression through the nuclear factor-κB signaling pathway in rat temporomandibular joint and condylar chondrocytes.pubmed:28337262

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