Overview

  • Product name
    Human Osteopontin ELISA Kit
    See all Osteopontin kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    serum 8 2.44%
    Inter-assay
    Sample n Mean SD CV%
    serum 3 3.26%
  • Sample type
    Cell culture supernatant, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    1.3 pg/ml
  • Range
    7.81 pg/ml - 500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Milk 105.3 102.6% - 109.1%
    Urine 107.39 104% - 109.4%
    Serum 97.92 96.8% - 98.7%
    Cell culture media 104.83 91.2% - 114%
    Heparin Plasma 96.19 93.3% - 99%
    EDTA Plasma 99.26 97.5% - 102%
    Citrate Plasma 94.56 90% - 99.6%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Osteopontin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Osteopontin protein in human serum, plasma, milk, urine and cell culture supernatant.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally,instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:
    Samples diluted in Sample Diluent 25BS –2.4 pg/mL
    Samples diluted in Sample Diluent NS –1.3pg/mL

  • Notes

    Osteopontin (OPN) is a 35kDa secreted adhesive glycophosphoprotein that belongs to the small integrin-binding ligand N-linked glycoprotein family (SIBLING) and it is expressed mainly in bone, teeth, kidney, epithelial lining tissues, activated T cells and macrophages and found in plasma and breast milk. It is processed by matrix metalloproteases 3 and 4 as well as thrombin, which generate two functional fragments: an N terminal (RGD) fragment that binds to integrin receptors and a C terminal fragment (containing heparin binding sites) that interacts with CD44. OPN can be present as a cytokine-like soluble protein or an immobilized extracellular matrix protein. OPN is important in multiple physiologic and pathologic processes including survival, adhesion, migration, angiogenesis, apoptosis, inflammation, macrophage migration, B cell proliferation, Ig production, wound healing and tumor metastasis.

    The proliferative and survival functions of OPN are mediated through αvβ3 integrin receptors as well as CD44 receptors. The downstream inhibition of apoptosis is mediated either by NF-kB or by the PI3K-Akt signaling pathway. High levels of OPN are a biomarker of poor prognosis in patients with breast cancer, multiple myeloma and prostate cancer. The aggressive nature of tumors with high expression of OPN has been linked to sustained activation of the epidermal growth factor receptor, increased migration of cancer cells in a αvβ3 integrin dependent fashion and regulation of HIF1alpha VEGF expression.

    OPN has also been shown to have a role in immune response. It is chemotactic for macrophages, dendritic cells and T cells. It influences both cell-mediated and humoral immunity as it has been shown to have Th1-cytokine functions as well as enhancing B lymphocytes proliferation and immunoglogulin production.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate (12 x 8 well strips)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab192143 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Ten individual healthy donors were evaluated for the presence of Osteopontin in serum using this assay. Results were interpolated from the standard curve in Sample Diluent NS and corrected by sample dilution (1:400). The mean level of Osteopontin is 79 ng/mL with a range of 14 – 174 ng/mL and a standard deviation of 45.5 ng/mL

Protocols

References

ab192143 has not yet been referenced specifically in any publications.

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