hh Human Amphiregulin Quantikine ELISA Kit DAR00: R&D Systems

Human Amphiregulin Quantikine ELISA Kit

(5 citations)
(2 Reviews)
  
  • 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), Cell Lysates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Saliva (50 uL), Urine (50 uL)
  • Sensitivity
    3.56 pg/mL
  • Assay Range
    15.6 - 1,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
  • Specificity
    Recombinant and natural human Amphiregulin
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Human Amphiregulin Immunoassay is a 4.5 hour solid phase ELISA designed to measure human Amphiregulin in cell culture supernates, cell lysates serum, plasma, saliva, and urine. It contains E. coli-derived recombinant human Amphiregulin and has been shown to accurately quantitate the recombinant factor. Results obtained with natural human Amphiregulin 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 natural human Amphiregulin.

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. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean10231463199.4308614
Standard Deviation5.6310.522.49.8520.528.8
CV%5.53.33.59.96.74.7

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 95 88-101
EDTA Plasma (n=4) 93 82-105
Heparin Plasma (n=4) 94 82-103
Saliva (n=4) 104 98-113
Serum (n=4) 95 85-103
Urine (n=4) 88 75-99
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Amphiregulin were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Amphiregulin Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Amphiregulin
Amphiregulin is a member of the EGF family of cytokines, which is comprised of at least ten proteins including EGF, TGF-alpha, HB-EGF, and the various heregulins. All of these cytokines are synthesized as transmembrane precursors and are characterized by the presence of one or several EGF structural units in their extracellular domain.

    • Entrez Gene IDs
      374 (Human); 11839 (Mouse);
    • Alternate Names
      Amphiregulin; AR; AREG; AREGB; Colorectum cell-derived growth factor; CRDGF; MGC13647; schwannoma-derived growth factor; SDGF;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    The conjugate must remain at 2-8 °C during use. Bring all other 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. 50 µL Assay Diluent
    4.   Add 50 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 cold Conjugate
    9.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 2 hours on the benchtop.
    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.

    5 Citations: Showing 1 - 5
    Filter your results:

    Species
    Sample Type
    1. ErbB activation signatures as potential biomarkers for anti-ErbB3 treatment in HNSCC
      Authors: D Alvarado, GF Ligon, JS Lillquist, SB Seibel, G Wallweber, VM Neumeister, DL Rimm, G McMahon, TM LaVallee
      PLoS ONE, 2017;12(7):e0181356.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Late acute graft versus host disease: a prospective analysis of clinical outcomes and circulating angiogenic factors
      Blood, 2016;0(0):.
      Species: Human
      Sample Type: Plasma
    3. Circulating hepatocyte growth factor is correlated with resistance to cetuximab in metastatic colorectal cancer.
      Authors: Yonesaka, Kimio, Satoh, Taroh, Ueda, Shinya, Yoshida, Takeshi, Takeda, Masayuki, Shimizu, Toshio, Okamoto, Isamu, Nishio, Kazuto, Tamura, Takao, Nakagawa, Kazuhiko
      Anticancer Res, 2015;35(3):1683-9.
      Species: Human
      Sample Type: Plasma
    4. Lipopolysaccharide (LPS)-Induced Biliary Epithelial Cell NRas Activation Requires Epidermal Growth Factor Receptor (EGFR).
      Authors: Trussoni C, Tabibian J, Splinter P, O'Hara S
      PLoS ONE, 2015;10(4):e0125793.
      Species: Human
      Sample Type: Cell Culture Supernates
    5. Endothelial cell-secreted EGF induces epithelial to mesenchymal transition and endows head and neck cancer cells with stem-like phenotype.
      Authors: Zhang Z, Dong Z, Lauxen I, Filho M, Nor J
      Cancer Res, 2014;74(10):2869-81.
      Species: Human
      Sample Type: Cell Culture Supernates
    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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    Cell Lysis Buffer 2

    ELISA 895347 2
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    Average Rating: 5 (Based on 2 reviews)

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      Excellent
      Anonymous 10/20/2017
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    Summary

    Sample TestedSUM-159PT human breast cancer cell line

    Other Experimental Details

    Other Experimental DetailsVery high sensitivity - no problem detecting amphiregulin released from human breast cancer cells.
     Human Amphiregulin Quantikine ELISA Kit DAR00
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    Excellent
      Anonymous 09/07/2017
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     Human Amphiregulin Quantikine ELISA Kit DAR00
    : Human Amphiregulin Quantikine ELISA Kit [DAR00]

    Summary

    Sample TestedEDTA Plasma

    Other Experimental Details

    Other Experimental DetailsThe Amphiregulin Quantikine ELISA kit performed as expected with low % CV's, and reproducible results.

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