Human Mesothelin Quantikine ELISA Kit

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DMSLN0
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Mesothelin ELISA Kit ELISA
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Product Details
Procedure
Citations (2)
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Human Mesothelin Quantikine ELISA Kit Summary

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.

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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 0.83 2.56 5.54 0.91 2.8 5.94
Standard Deviation 0.034 0.082 0.193 0.057 0.13 0.36
CV% 4 3.2 3.5 6.2 4.6 6.1

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 ELISA Linearity

Data Examples

Human Mesothelin ELISA Standard Curve

Product Datasheets

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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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 for Human Mesothelin Quantikine ELISA Kit

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.

2 Citations: Showing 1 - 2
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  1. Assessment of potential predictors of calretinin and mesothelin to improve the diagnostic performance to detect malignant mesothelioma: results from a population-based cohort study
    Authors: S Casjens, DG Weber, G Johnen, I Raiko, D Taeger, C Meinig, S Moebus, KH Jöckel, T Brüning, B Pesch
    BMJ Open, 2017;7(10):e017104.
    Species: Human
    Sample Types: Plasma
  2. 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 Types: Plasma

FAQs

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Human Mesothelin Quantikine ELISA Kit
By Anonymous on 08/17/2018
Sample Tested: Cell Culture Supernates