Overview

  • Product name

    Mouse Mesothelin ELISA Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 8.3%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 9.8%
  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    14 pg/ml
  • Range

    125 pg/ml - 8000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 119.6 115.6% - 122.8%
    Cell culture media 88.5 85.6% - 90%
    EDTA Plasma 116.3 114.4% - 118.8%
    Citrate Plasma 112.1 108.2% - 114.6%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rat, Rabbit, Hamster, Dog, Human, Pig
  • Product overview

    Abcam’s Mesothelin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Mesothelin protein in mouse serum, plasma, cell culture supernatants and, cell and tissue extract samples. (Genecards: Msln)


    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 NS: 41 pg/mL


    Samples diluted in Sample Diluent 25BS: 70 pg/mL


    Samples diluted in 1X Cell Extraction Buffer PTR: 14 pg/mL

  • Notes

    Mesothelin is expressed as a much larger, 70 kDa precursor, which is cleaved into two functional proteins, Megakaryocyte potentiating factor and Mesothelin. Mesothelin is a glycosylated cell-surface protein, anchored to the plasma membrane via its GPI anchor. Membrane-anchored forms of Mesothelin may play a role in cellular adhesion.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse Mesothelin Capture Antibody 1 x 600µl
    10X Mouse Mesothelin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI 1 x 6ml
    Mouse Mesothelin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent 25BS 1 x 20ml
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Cellular localization

    Secreted and Cell membrane. Golgi apparatus.
  • Alternative names

    • CAK1 antigen
    • Megakaryocyte potentiating factor
    • MPF
    • MSLN
    • Pre pro megakaryocyte potentiating factor
    • SMRP
    • Soluble MPF mesothelin related protein
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab204528 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 curve and corrected for sample dilution. Note that 1X Diluted samples were pre-diluted to 50%. The interpolated, dilution factor-corrected values are plotted in pg of Mesothelin per mL of neat sample (mean +/- SD, n=2).

  • The concentrations of Mesothelin were measured in duplicates, interpolated from the Mesothelin standard curve and corrected for sample dilution. Note that 1X Diluted C2C12 cell extract samples were pre-diluted to 125 µg/mL. Note that 1X Diluted NIH/3T3 cell extract samples were pre-diluted to 250 µg/mL. The interpolated, dilution factor-corrected values are plotted in pg of Mesothelin per mg of extract (mean +/- SD, n=2).

Protocols

References

ab204528 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Answer

Thank you for getting in touch.




Both capture and detector antibodies were generated using Mesothelin cleaved form (amino acids 298-600 of the precursor protein, http://www.uniprot.org/uniprot/Q61468). Therefore, this kit cannot measure Megakaryocyte-potentiating factor (amino acids 36-288).



The reactivity with the precursor Mesothelin (amino acids 36-600) was not tested.



I am happy to answer any further questions you have.

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