ELISA Kit for Anti-Osteopontin Antibody (Anti-OPN)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 90-101 96
EDTA plasma(n=5) 82-98 94
heparin plasma(n=5) 97-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Anti-Osteopontin Antibody (Anti-OPN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Anti-Osteopontin Antibody (Anti-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 Anti-Osteopontin Antibody (Anti-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) 92-101% 80-101% 97-105% 91-99%
EDTA plasma(n=5) 80-98% 99-105% 79-98% 80-101%
heparin plasma(n=5) 99-105% 93-105% 96-105% 93-101%

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
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 5 times;
5. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
6. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Journal of Biomedical Materials Research Part A Proliferation and osteogenesis of immortalized bone marrow-derived mesenchymal stem cells in porous polylactic glycolic acid scaffolds under perfusion culture PubMed: 19280635
Osteoarthritis and Cartilage Elevated osteopontin level of synovial fluid and articular cartilage is associated with disease severity in knee osteoarthritis patients PubMed: 19747583
The Journal of Rheumatology Thrombin-cleaved osteopontin levels in synovial fluid correlate with disease severity of knee osteoarthritis. Jrheum: source
International Journal of Pharmaceutics Suppression of tumor growth in xenograft model mice by small interfering RNA targeting osteopontin delivery using biocompatible poly(amino ester) ScienceDirect: S0378517312003742
European Cells and matreials The use of non-viral gene vectors for bioactive poly-(D, L-lactide) implant surfaces in bone tissue engineering Ecmjournal: Source
Journal of Equine Veterinary Science Influence of Seminal Plasma Antioxidants and Osteopontin on Fertility of the Arabian Horse ScienceDirect: Source
Basic &amp; Clinical Pharmacology &amp; Toxicology Treatment with Carnitine Enhances Bone Fracture Healing under Osteoporotic and/or Inflammatory Conditions Wiley:Source
Journal of Cellular Biochemistry Tension Force-Induced ATP Promotes Osteogenesis Through P2X7 Receptor in Osteoblasts Pubmed:24905552
Naunyn-Schmiedeberg&#039;s Archives of Pharmacology Treatment with α-lipoic acid enhances the bone healing after femoral fracture model of rats Pubmed:25038619
J Endocrinol Invest.? The interplay among iron metabolism, endothelium and inflammatory cascade in dysmetabolic disorders Pubmed:25245337
FEBS Letters Low-intensity pulsed ultrasound-induced ATP increases bone formation via the P2X7 receptor in osteoblast-like MC3T3-E1 cells Pubmed:25542352
Gene Effects of magnesium degradation products on mesenchymal stem cell fate and osteoblastogenesis PubMed: 26283150
Medicine (Baltimore) An Attempt to Evaluate Selected Aspects of “Bone–Fat Axis” Function in Healthy Individuals and Patients With Pancreatic Cancer PubMed: 26266370
Journal of Biomaterials and Tissue Engineering Osteogenic Differentiation of Canine Adipose Stem Cells Cultured in Alginate-Fibrin-Based Hydrogel Content: Asp
Primasari Osteopontin level correlates negatively with tumor shrinkage in neoadjuvant chemoradiation of locally advanced rectal cancer Index.Php: Amor
Int J Biol Sci Defective Initiation of Liver Regeneration in Osteopontin-Deficient Mice after Partial Hepatectomy due to Insufficient Activation of IL-6/Stat3 Pathway PubMed: 26327817
Journal of Kunming Medical University / Kunming Yike Daxue Xuebao Perioperative 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 journal Stenotic Bicuspid and Tricuspid Aortic Valves - Micro-Computed Tomography and Biological Indices of Calcification. pubmed:28344201
Journal of Cardiovascular Electrophysiology Effect of Cardiac Resynchronization Therapy on Myocardial Fibrosis and Relevant Cytokines in a Canine Model With Experimental Heart Failure. pubmed:28127817
American Journal of Translational Research Osteopontin stimulates matrix metalloproteinase expression through the nuclear factor-κB signaling pathway in rat temporomandibular joint and condylar chondrocytes. pubmed:28337262
Diabetes Care Targeted Microbiome Intervention by Microencapsulated Delayed-Release Niacin Beneficially Affects Insulin Sensitivity in Humans pubmed:29212824
Biomedicine & pharmacotherapy Icaritin induces MC3T3-E1 subclone14 cell differentiation through estrogen receptor-mediated ERK1/2 and p38 signaling activation. pubmed:28742995
Journal of Alzheimers Disease Urine-Based Biomarkers for Alzheimer's Disease Identified Through Coupling Computational and Experimental Methods Pubmed:30040720
Indian Journal of Nephrology Osteopontin, cardiovascular risk factors and carotid intima-Media thickness in chronic kidney disease id:233806
Carbon In Situ Self-Assembly of Graphene Oxide/Polydopamine/Sr2+ Nanosheets on Titanium Surfaces for Enhanced Osteogenic Differentiation of Mesenchymal Stem Cells Doi: 10.1016/j.carbon.2018.10.081
current eye research Detection of proteins associated with extracellular matrix regulation in the aqueous humour of patients with primary glaucoma. Pubmed: 30994369
Journal of Proteomics Identification of urinary candidate biomarkers of cisplatin-induced nephrotoxicity in patients with carcinoma Pubmed: 31629959
Acta Biomaterialia Inflammatory response to magnesium-based biodegradable implant materials Pubmed: 31610341
Journal of Clinical Medicine Calcium Kidney Stones are Associated with Increased Risk of Carotid Atherosclerosis: the Link between Urinary Stone Risks, Carotid Intima-Media Thickness, and … Pubmed: 32182704
EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES Febuxostat attenuates vascular calcification induced by vitamin D3 plus nicotine in rats Pubmed: 33010420
[ENGINEERING OF BIOMATERIALS Osteoconductive potential of PLGA/bioglass composite biomaterials
Biomolecules Structural Constraint of Osteopontin Facilitates Efficient Binding to CD44 34070790
Structural Changes and Detection of Liver Fibrosis¨CRelated Protein Levels in Coculture of Alveolar Echinococcosis-Protoscoleces and Human Hepatic Stellate cells
Parasit Vectors Structural changes and expression of hepatic fibrosis-related proteins in coculture of Echinococcus multilocularis protoscoleces and human hepatic stellate … 34857049
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