Multiplex Assay Kit for Osteopontin (OPN) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Osteopontin (OPN) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Osteopontin (OPN) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 78-101 82
EDTA plasma(n=5) 89-96 93
heparin plasma(n=5) 81-101 90
sodium citrate plasma(n=5) 83-96 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Osteopontin (OPN) ,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 Osteopontin (OPN) ,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 Osteopontin (OPN) ,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) 79-91% 87-101% 80-103% 80-93%
EDTA plasma(n=5) 92-103% 97-105% 95-102% 89-98%
heparin plasma(n=5) 82-101% 79-95% 78-92% 88-99%
sodium citrate plasma(n=5) 95-105% 82-101% 80-103% 82-92%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:OPN) 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.

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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
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[ENGINEERING OF BIOMATERIALS Osteoconductive potential of PLGA/bioglass composite biomaterials
Biomolecules Structural Constraint of Osteopontin Facilitates Efficient Binding to CD44 34070790
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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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Catalog No. Related products for research use of Sus scrofa; Porcine (Pig) Organism species Applications (RESEARCH USE ONLY!)
RPA899Po01 Recombinant Osteopontin (OPN) Positive Control; Immunogen; SDS-PAGE; WB.
PAA899Po01 Polyclonal Antibody to Osteopontin (OPN) WB; IHC; ICC; IP.
MAA899Po21 Monoclonal Antibody to Osteopontin (OPN) WB; IHC; ICC; IP.
MAA899Po22 Monoclonal Antibody to Osteopontin (OPN) WB; IHC; ICC; IP.
MAA899Po23 Monoclonal Antibody to Osteopontin (OPN) WB; IHC; ICC; IP.
MAA899Po24 Monoclonal Antibody to Osteopontin (OPN) WB; IHC; ICC; IP.
SEA899Po ELISA Kit for Osteopontin (OPN) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA899Po Multiplex Assay Kit for Osteopontin (OPN) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.