Human Osteopontin,OPN ELISA Kit

Code CSB-E08392h
Size 96T,5×96T,10×96T
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Product Details

Target Name
secreted phosphoprotein 1
Alternative Names
BNSP ELISA Kit; Bone sialoprotein 1 ELISA Kit; Bone sialoprotein I ELISA Kit; BSP I ELISA Kit; BSPI ELISA Kit; Early T lymphocyte activation 1 ELISA Kit; ETA 1 ELISA Kit; ETA1 ELISA Kit; MGC110940 ELISA Kit; Nephropontin ELISA Kit; OPN ELISA Kit; Osteopontin ELISA Kit; osteopontin/immunoglobulin alpha 1 heavy chain constant region fusion protein ELISA Kit; OSTP_HUMAN ELISA Kit; PSEC0156 ELISA Kit; secreted phosphoprotein 1 (osteopontin bone sialoprotein I early T lymphocyte activation 1) ELISA Kit; secreted phosphoprotein 1 (osteopontin, bone sialoprotein I, early T-lymphocyte activation 1) ELISA Kit; Secreted phosphoprotein 1 ELISA Kit; SPP 1 ELISA Kit; SPP-1 ELISA Kit; SPP1 ELISA Kit; SPP1/CALPHA1 fusion ELISA Kit; Urinary stone protein ELISA Kit; Uropontin ELISA Kit
Abbreviation
OPN
Uniprot No.
Species
Homo sapiens (Human)
Sample Types
serum, plasma, urine, tissue homogenates
Detection Range
0.78 ng/mL-50 ng/mL
Sensitivity
0.195 ng/mL
Assay Time
1-5h
Sample Volume
50-100ul
Detection Wavelength
450 nm
Research Area
Cancer
Assay Principle
quantitative
Measurement
Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human OPN in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:5 Average % 94  
Range % 88-98  
1:10 Average % 100  
Range % 94-104  
1:20 Average % 93  
Range % 88-97  
1:40 Average % 99  
Range % 93-104  
Recovery
The recovery of human OPN spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 95 81-97  
EDTA plasma (n=4) 102 97-106  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
50 2.418 2.333 2.376 2.271  
25 1.684 1.706 1.695 1.590  
12.5 1.175 1.152 1.164 1.059  
6.25 0.654 0.664 0.659 0.554  
3.12 0.383 0.361 0.372 0.267  
1.56 0.245 0.232 0.239 0.134  
0.78 0.176 0.185 0.181 0.076  
0 0.102 0.108 0.105    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human OPN antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled OPN antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
Storage
Store at 2-8°C. Please refer to protocol.
Lead Time
3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx
Description

The Human Osteopontin (OPN) ELISA Kit is a powerful tool for researchers in the field of cancer. This quantitative assay measures levels of OPN, a protein involved in the regulation of bone remodeling and implicated in various diseases including cancer(tumor),bone metabolism, immunity, growth and development.

With a detection range of 0.78 ng/mL-50 ng/mL and a sensitivity of 0.195 ng/mL, this ELISA kit can detect low levels of OPN with high accuracy. It can be used with a variety of sample types, including serum, plasma, urine, and tissue homogenates, making it a versatile tool for research.

The assay principle is based on the sandwich method, where a monoclonal antibody specific to human OPN is pre-coated onto a microplate. The sample is added, and any OPN present in the sample binds to the antibody. A biotinylated polyclonal antibody specific to human OPN is then added, which binds to the OPN-antibody complex. Finally, streptavidin conjugated to horseradish peroxidase (HRP) is added, which binds to the biotinylated antibody. The HRP produces a colorimetric reaction in the presence of a substrate, which can be measured at a wavelength of 450 nm.

The assay can be completed in as little as 1-5 hours and requires a sample volume of only 50-100ul, minimizing the amount of sample required for analysis. The kit includes all necessary components and can be used with standard laboratory equipment.

With 12 citations, this kit has been validated by the scientific community and is a trusted tool for cancer research. So, whether you are studying the role of OPN in cancer progression or investigating potential therapeutic targets, this Human Osteopontin (OPN) ELISA Kit is an essential addition to your research arsenal.

Citations

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Target Background

Function
(From Uniprot)
Major non-collagenous bone protein that binds tightly to hydroxyapatite. Appears to form an integral part of the mineralized matrix. Probably important to cell-matrix interaction.; Acts as a cytokine involved in enhancing production of interferon-gamma and interleukin-12 and reducing production of interleukin-10 and is essential in the pathway that leads to type I immunity.
Gene References into Functions
  1. Plasma OPN levels were associated with the presence and severity of diabetic retinopathy in Asians with type 2 diabetes. PMID: 29463954
  2. Study demonstrated that osteopontin exerts an important role in the monocytes/macrophage phenotypic differentiation from hypertensive patients with vascular calcification. PMID: 28091516
  3. The expression of OPN was high in endometrium in secretory phase and in vitro decidualized endometrial stromal cells. Further analysis confirmed that OPN expression was upregulated by cAMP and C/EBPbeta signal pathway, while downregulated by miR181b. Increased OPN expression could promote the expression of decidualization-related and angiogenesis-related genes PMID: 29420252
  4. Results of the study showed that low urinary OPN levels were correlated with increased kidney stone risk, and dietary habits can affect urinary OPN level PMID: 30114399
  5. The serum levels of cystatin C and urine NGAL, urine OPN can be used as a good marker for assessing the renal effect of obesity which can lead end stage renal disease in pediatric population. PMID: 30035656
  6. novel 9175th- (exon 7) position polymorphism of OPN and rs17524488 were related to susceptibility to ankylosing spondylitis in a Chinese population PMID: 29581970
  7. Over-expressed and hypo-methylated SPP1 gene is associated with hepatocellular carcinoma. PMID: 27760737
  8. Serum osteopontin concentrations were increased in acute pancreatitis (AP) compared with follow-up 3 months after discharge. PMID: 29424811
  9. Our data provide evidence that leptin-mediated OPN upregulation promote TH2 inflammation in AR and this process is achieved through the alpha4 integrin and PI3K/AKT signaling pathways. PMID: 29885866
  10. miR-129-5p level was decreased in fibrotic liver of human, and reduced by rOPN treatment. In contrast, miR-129-5p was induced in HSCs transfected by OPN siRNA. These data suggested that OPN induces Col 1 expression via suppression of miR-129-5p in hepatic stellate cells. PMID: 29196165
  11. High circulating osteopontin levels predict major adverse cardiovascular events in patients with severe carotid artery stenosis. PMID: 29317141
  12. High SPP1 Expression is associated with Hepatocellular Carcinomas. PMID: 29693976
  13. Serum OPN and DKK1 levels of hepatocellular carcinoma patients could be considered as novel biomarkers showing prognostic significance after hepatectomy based on long-term survival data PMID: 29753515
  14. Upregulation of ADIPOR1 and SPP1, among the adipokine gene family, in cancer tissue is associated with poor survival in CRC, suggesting a potential mechanism linking obesity and colorectal cancer. PMID: 29761507
  15. Review/Meta-analysis: report a significantly higher circulating OPN level in systemic lupus erythematosus patients, a trend of positive correlation between OPN levels and SLE activity, and a significant association between OPN 1239 C/A and 9250 C/T polymorphisms, and SLE development. PMID: 27307447
  16. OPN is not only the pivotal protein controlling fibrosis, but may also serve as a biomarker associated with prognosis. PMID: 29120858
  17. Our data show that the ability of AML cells to constitutively release high levels of osteopontin is associated with a favorable prognosis, and this is different from previous studies of osteopontin mRNA levels. PMID: 27739925
  18. First report of OPN cleavage in THP-1 macrophages after PMA (phorbol 12-myristate 13-acetate) stimulation, and of enhanced cleavage induced by BCG infection (bovine tuberculosis). PMID: 29385060
  19. The results suggest that OPN, MMP9 and S100A8 all play a significant role in bladder cancer progression and are potential prognostic markers and therapeutic targets in bladder cancer PMID: 29209142
  20. We demonstrated an important effect of the SPP1 rs4754 on subclinical markers of carotid atherosclerosis in subjects with T2DM; however, as demonstrated by the multiple linear regression analysis, neither rs4754 nor rs28357094 had an important impact on the progression of subclinical markers of carotid atherosclerosis in subjects with T2DM. PMID: 28990744
  21. these results showed that expression of tear OPN protein by perennial allergic conjunctivitis patients increased significantly compared with controls or seasonal allergic conjunctivitis patients out of the pollen season PMID: 29279263
  22. The study revealed an enhanced sensitivity of aortic valve interstitial cells to osteogenic inductors in aortic stenosis patients, which indicates probable implication of OPN, OPG, and BMP2 genes in pathogenesis of aortic valve calcification. PMID: 29308559
  23. Carriers of the risky genotype or haplotype also had increased gastric OPN expression (p = 0.038) and inflammation (p = 0.007). SPP1 polymorphisms predispose to intestinal metaplasia development in H. pylori infected males. PMID: 28685609
  24. Respiratory syncytial virus infection leads to increased OPN expression.IL-1beta is involved in the regulation of OPN levels during respiratory syncytial virus infection. PMID: 29677209
  25. plasma level increases in pregnancy independently of asthma PMID: 29200898
  26. Methylglyoxal-bis-guanylhydrazone may have potential therapeutic utility in reducing or normalizing OPN levels and regulating monocyte activation in diseases that involve chronic inflammation. PMID: 29538412
  27. These results might be considered a useful starting point to validate OPN as a prognostic and diagnostic marker of hepatocellular carcinoma. PMID: 28711012
  28. study demonstrates that the OPN gene SNP rs1126616 is statistically associated with cerebral palsy. PMID: 27114095
  29. LAMP3 promotes the invasion of osteosarcoma cells via SPP1 signaling. PMID: 28849219
  30. Extended liver surgery is the only potentially curative treatment for patients with cholangiocarcinoma (CCA/biliary cancer), but it is currently unclear which patients benefit most from surgery. Detecting serum levels of osteopontin - a specific secreted glycoprotein involved in multiple diseases - in CCA patients might help to identify those patients that particularly benefit from tumour resection. PMID: 28668580
  31. MALAT1 could directly bind to miR-127-5p to inhibit its expression, so as to rescue OPN expression and promote chondrocyte proliferation through PI3K/Akt pathway. PMID: 28590075
  32. A linear negative correlation between serum OPN and total bone mineral density (BMD) of Type I diabetes mellitus patients compared to control group was noticed, indicating that the amount of serum OPN could be effective on BMD and a good predicting factor for osteoporosis PMID: 28499311
  33. OPN regulates CYP7A1 levels and the metabolic fate of liver acetyl-CoA as a result of CHOL and PC metabolism interplay PMID: 28754826
  34. these findings demonstrated that OPN and VEGF were overproduced in nasal polyps and that OPN induced VEGF production, which indicated that OPN-VEGF axis might contribute to angiogenesis in nasal polyps PMID: 28716167
  35. Osteopontin and CD44 play important roles in the development and progression of meningioma and can be used as prognostic markers for tumor recurrence and progression as well as therapeutic targets for the development of new drugs. PMID: 29504367
  36. OPN play a role in pathogenesis of neurodegenerative diseases or in neuroprotection by regulating the activation and function of microglia. PMID: 28698867
  37. osteopontin has a role in DNA repair and impact on human glioblastoma radiosensitivity PMID: 27563812
  38. SPP1 promoted the metastasis of CRC. PMID: 28531945
  39. Results show that osteopontin is positively associated with hepatocellular carcinoma (HCC) metastasis, and that OPN can activate CCR1 expression through the PI3K/AKT/HIF-1a signaling pathway to promote HCC progression and metastasis. PMID: 29285854
  40. This review presents the current knowledge on the expression profiles of OPN and its main splice variants in human cancers, as well as the potential implications in patient outcome. [review] PMID: 28440483
  41. Strong correlations between the expression of type I, II, IV collagen and osteopontin and the clinical stage of tympanosclerosis indicate the involvement of these proteins in excessive fibrosis and pathological remodeling of the tympanic membrane. PMID: 29068597
  42. result of this study confirms that increased levels of OPN exist in CSF and peripheral blood of MS patients and strengthens the evidence regarding the clinical utility of OPN as a promising and validated biomarker for MS PMID: 29346446
  43. OPN overexpression was correlated with poor overall survival and clinical features reflecting high aggressiveness in patients with gastric cancer. PMID: 27626167
  44. ITGbeta3 and CD44 expression levels determine whether OPN-a inhibits or enhances growth in lung cancer cells. PMID: 27487131
  45. Study identified that miR-127-5p targets the 3' UTR of osteoarthritis (OA) mRNA to down-regulate the expression of OPN. In OA, the down-expressed miR-127-5p allows the expression of OPN, which mediates the establishment and development of OA. PMID: 27126955
  46. Our study shows a strong effect of the rs28357094 G allele in increasing osteopontin expression in the presence of deflazacort, and adds to the evidence that rs28357094 polymorphism may predict response to glucocorticoids in Duchenne muscular dystrophy. PMID: 28595270
  47. we described the expression pattern of osteopontin splice variants in papillary thyroid carcinoma samples and the key role of osteopontin-a expression on activating tumor progression features PMID: 27409830
  48. In Osteoarthritis tissues, OPN mRNA, and NEAT1 expression was upregulated, whereas miR-181c expression was downregulated, indicating that targeting NEAT1 to rescue miR-181c expression so as to inhibit OPN expression and synoviocyte proliferation PMID: 28379604
  49. IL-6 and sIL-6R induced by IL-1beta may trigger IL-6 trans-signaling, contributing to the upregulation of OPN in THP-1 macrophages. Macrophages may be used as a source of IL-6 and sIL-6R and evoke IL-6 trans-signaling. PMID: 27863335
  50. OPN is a key mediator of intracerebral tumor growth, invasion, and dissemination in central nervous system lymphoma, and these effects depend upon activation of NF-kappaB. PMID: 27050077

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Subcellular Location
Secreted.
Protein Families
Osteopontin family
Tissue Specificity
Bone. Found in plasma.
Database Links

HGNC: 11255

OMIM: 166490

KEGG: hsa:6696

STRING: 9606.ENSP00000378517

UniGene: Hs.313

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