Overview

  • Product name

    Human Mesothelin ELISA Kit
    See all Mesothelin kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 3.7%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 3.3%
  • Sample type

    Cell culture supernatant, Saliva, Urine, Serum, Heparin Plasma, EDTA Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    132.5 pg/ml
  • Range

    0.39 ng/ml - 25 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 95 92% - 97%
    Saliva 104 100% - 107%
    Urine 87 85% - 89%
    Serum 89 84% - 95%
    Heparin Plasma 84 81% - 87%
    EDTA Plasma 98 88% - 111%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

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


    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.

  • Notes

    Mesothelin is a 40 kDa GPI-anchored cell surface protein found on mesothelial cells and overexpressed in several types of cancer. The gene that encodes mesothelin is spliced into two different gene products, mesothelin and megakaryocyte potentiation factor. Mesothelin is involved in cellular adhesion, but is also secreted and normally found in several biological fluids.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human Mesothelin Capture Antibody 1 x 600µl
    10X Human Mesothelin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BI 1 x 6ml
    Human Mesothelin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Membrane-anchored forms may play a role in cellular adhesion.
    Megakaryocyte-potentiating factor (MPF) potentiates megakaryocyte colony formation in vitro.
  • Tissue specificity

    Expressed in lung. Expressed at low levels in heart, placenta and kidney. Expressed in mesothelial cells. Highly expressed in mesotheliomas, ovarian cancers, and some squamous cell carcinomas (at protein level).
  • Involvement in disease

    Note=Antibodies against MSLN are detected in patients with mesothelioma and ovarian cancer.
  • Sequence similarities

    Belongs to the mesothelin family.
  • Post-translational
    modifications

    Both MPF and the cleaved form of mesothelin are N-glycosylated.
    Proteolytically cleaved by a furin-like convertase to generate megakaryocyte-potentiating factor (MPF), and the cleaved form of mesothelin.
  • Cellular localization

    Secreted and Cell membrane. Golgi apparatus.
  • Information by UniProt
  • Alternative names

    • CAK 1
    • CAK1
    • CAK1 antigen
    • cleaved form
    • Megakaryocyte potentiating factor
    • Mesothelin
    • Mesothelin isoform 1 precursor
    • MPF
    • Msln
    • MSLN_HUMAN
    • Pre pro megakaryocyte potentiating factor
    • Pre-pro-megakaryocyte-potentiating factor
    • SMR
    • SMRP
    • Soluble MPF mesothelin related protein
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab216168 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

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

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

  • The concentrations of Mesothelin were measured in duplicates, interpolated from the Mesothelin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (heparin) 37.5%, plasma (EDTA) 37.5%, urine 50% and saliva 5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Mesothelin concentration was determined to be 24.50 ng/mL in serum, 17.17 ng/mL in plasma (heparin), 24.54 ng/mL in plasma (EDTA), 12.37 ng/mL in urine and 277.2 ng/mL in saliva.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Mesothelin concentration was determined to be 25.79 ng/mL with a range of 6.86 – 60.93 ng/mL.

Protocols

References

ab216168 has not yet been referenced specifically in any publications.

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