Human Amphiregulin Quantikine ELISA Kit

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Control Products Available
Human Amphiregulin ELISA Standard Curve
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Product Details
Procedure
Citations (8)
FAQs
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Reviews (5)

Human Amphiregulin 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), 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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 102 314 631 99.4 308 614
Standard Deviation 5.63 10.5 22.4 9.85 20.5 28.8
CV% 5.5 3.3 3.5 9.9 6.7 4.7

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 ELISA Linearity

Data Examples

Human Amphiregulin ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Amphiregulin

Amphiregulin (AR) is an EGF family growth factor that is released as a soluble protein following proteolytic cleavage of its transmembrane precursor. Amphiregulin is expressed by numerous carcinoma cell lines and epithelial cells of the colon, stomach, breast, ovary and kidney. It acts through ErbB family of receptor to stimulate the proliferation of keratinocytes, mammary epithelial cells, fibroblasts, astrocytes and glial cells. Amphiregulin can also inhibit the growth of certain tumor cells.

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
⚠ 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.

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 for Human Amphiregulin 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.

8 Citations: Showing 1 - 8
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  1. Targeting amphiregulin (AREG) derived from senescent stromal cells diminishes cancer resistance and averts programmed cell death 1 ligand (PD-L1)-mediated immunosuppression
    Authors: Q Xu, Q Long, D Zhu, D Fu, B Zhang, L Han, M Qian, J Guo, J Xu, L Cao, YE Chin, JP Coppé, EW Lam, J Campisi, Y Sun
    Aging Cell, 2019;0(0):e13027.
    Species: Human
    Sample Types: Serum
  2. Combinations of Bevacizumab and Erlotinib show activity in colorectal cancer independent of RAS status
    Authors: P Mésange, A Bouygues, N Ferrand, M Sabbah, AE Escargueil, A Savina, B Chibaudel, C Tournigand, T Andre, A de Gramont, AK Larsen
    Clin. Cancer Res., 2018;0(0):.
    Species: Human
    Sample Types: Whole Cells
  3. Yes-associated protein promotes the abnormal proliferation of psoriatic keratinocytes via an amphiregulin dependent pathway
    Authors: J Jia, C Li, J Yang, X Wang, R Li, S Luo, Z Li, J Liu, Z Liu, Y Zheng
    Sci Rep, 2018;8(1):14513.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. 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 Types: Cell Culture Supernates
  5. Late acute graft versus host disease: a prospective analysis of clinical outcomes and circulating angiogenic factors
    Blood, 2016;0(0):.
    Species: Human
    Sample Types: Plasma
  6. 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 Types: Cell Culture Supernates
  7. 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 Types: Plasma
  8. 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 Types: Cell Culture Supernates

FAQs

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Reviews for Human Amphiregulin Quantikine ELISA Kit

Average Rating: 5 (Based on 5 Reviews)

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Human Amphiregulin Quantikine ELISA Kit
By Anonymous on 04/18/2018
Application: Sample Tested: colorectal adenocarcinoma cell line

Human Amphiregulin Quantikine ELISA Kit
By Anonymous on 04/05/2018
Application: Sample Tested: human pancreatic cell line

Human Amphiregulin Quantikine ELISA Kit
By Anonymous on 01/11/2018
Application: Sample Tested: Serum

Human Amphiregulin Quantikine ELISA Kit
By Anonymous on 10/20/2017
Application: Sample Tested: SUM-159PT human breast cancer cell line

Very high sensitivity - no problem detecting amphiregulin released from human breast cancer cells.


Human Amphiregulin Quantikine ELISA Kit
By Anonymous on 09/07/2017
Application: Sample Tested: EDTA Plasma

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