Human Mesothelin Quantikine ELISA Kit

(1 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (10 uL), Urine (50 uL)
  • Sensitivity
    0.022 ng/mL
  • Assay Range
    0.2 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
  • Specificity
    Natural and recombinant human Mesothelin
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC225, Quantikine Immunoassay Control Set 717 for Human Mesothelin - Please Inquire
Product Summary
The Quantikine Human Mesothelin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Mesothelin in cell culture supernates, serum, plasma, saliva, and urine. It contains NS0-expressed recombinant human Mesothelin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Mesothelin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring Mesothelin.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean0.832.565.540.912.85.94
Standard Deviation0.0340.0820.1930.0570.130.36
CV%43.23.56.24.66.1

Recovery

The recovery of Mesothelin spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=8) 104 91-113
EDTA Plasma (n=4) 97 88-109
Heparin Plasma (n=4) 97 86-112
Saliva (n=4) 102 91-112
Serum (n=4) 99 92-114
Urine (n=4) 103 86-115
Linearity
To assess the linearity of the assay, samples containing high concentrations of Mesothelin were serially diluted Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Mesothelin Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Mesothelin
Mesothelin, also known as CAK1 and ERC, is derived from a precursor that also includes Megakaryocyte Potentiating Factor (MPF). Following cleavage of the precursor, Mesothelin remains attached to the cell surface via a GPI linkage. Mesothelin is normally expressed on mesothelial cells in the pleura, pericardium, and peritoneum as well as in the developing and postnatal pancreas. It is upregulated in mesotheliomas and a range of carcinomas and adenomas. Mesothelin promotes tumor cell proliferation, migration, anchorage-independent growth, and tumor progression.
    • Entrez Gene IDs
      10232 (Human); 56047 (Mouse); 60333 (Rat);
    • Alternate Names
      CAK1 antigen; CAK1; megakaryocyte potentiating factor; Mesothelin; MPF; MPFSMRP; MSLN; Pre-pro-megakaryocyte-potentiating factor; SMR; soluble MPF mesothelin related protein;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    1 Citations: Showing 1 - 1

    1. Expansion of anti-mesothelin specific CD4+ and CD8+ T cell responses in patients with pancreatic carcinoma.
      Authors: Chen Y, Ayaru L, Mathew S, Morris E, Pereira S, Behboudi S
      PLoS ONE, 2014;9(2):e88133.
      Species: Human
      Sample Type: Plasma
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 717 for Human Mesothelin

    Ctrl QC225
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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