Human Amphiregulin Quantikine ELISA Kit

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Human Amphiregulin ELISA Standard Curve
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Citations (18)
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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 (pg/mL) 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

Scientific Data

Human Amphiregulin ELISA Standard Curve

Product Datasheets

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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
&#9888; 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.

18 Citations: Showing 1 - 10
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  1. Identifying immune signatures of sepsis to increase diagnostic accuracy in very preterm babies
    Authors: Das, A;Ariyakumar, G;Gupta, N;Kamdar, S;Barugahare, A;Deveson-Lucas, D;Gee, S;Costeloe, K;Davey, MS;Fleming, P;Gibbons, DL;
    Nature communications
    Species: Human
    Sample Types: Plasma
  2. EGF, TGF-? and Amphiregulin Differently Regulate Endometrium-Derived Mesenchymal Stromal/Stem Cells
    Authors: Kamentseva, RS;Kharchenko, MV;Gabdrahmanova, GV;Kotov, MA;Kosheverova, VV;Kornilova, ES;
    International journal of molecular sciences
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Luteinizing hormone stimulates the expression of amphiregulin in human theca cells
    Authors: Y Liu, Y Zhong, X Shen, X Guo, R Wu, T Yang, M Chen
    Journal of ovarian research, 2022-12-07;15(1):129.
    Species: Human
    Sample Types: Cell Lysates
  4. iRhom2 regulates ERBB signalling to promote KRAS-driven tumour growth of lung cancer cells
    Authors: B Sieber, F Lu, SM Stribbling, AG Grieve, AJ Ryan, M Freeman
    Journal of Cell Science, 2022-09-08;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Amphiregulin can predict treatment resistance to palliative first-line cetuximab plus FOLFIRI chemotherapy in patients with RAS wild-type metastatic colorectal cancer
    Authors: SA Kim, H Park, KJ Kim, JW Kim, JH Sung, M Nam, JH Lee, EH Jung, KJ Suh, JY Lee, SH Kim, JO Lee, JW Kim, YJ Kim, JH Kim, SM Bang, JS Lee, KW Lee
    Scientific Reports, 2021-12-10;11(1):23803.
    Species: Human
    Sample Types: Plasma
  6. Unraveling the differential impact of PAHs and dioxin-like compounds on AKR1C3 reveals the EGFR extracellular domain as a critical determinant of the AHR response
    Authors: C Vogeley, NC Sondermann, S Woeste, AA Momin, V Gilardino, F Hartung, M Heinen, SK Maa beta, M Mescher, M Pollet, KM Rolfes, CFA Vogel, A Rossi, D Lang, ST Arold, M Nakamura, T Haarmann-S
    Environment international, 2021-11-20;158(0):106989.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Secreted matrix metalloproteinase-14 is a predictor for antifibrotic effect of IC-2-engineered mesenchymal stem cell sheets on liver fibrosis in mice
    Authors: K Fukushima, N Itaba, Y Kono, S Okazaki, S Enokida, N Kuranobu, J Murakami, M Enokida, H Nagashima, S Kanzaki, N Namba, G Shiota
    Regenerative Therapy, 2021-08-26;18(0):292-301.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Pulmonary fibrosis distal airway epithelia are dynamically and structurally dysfunctional
    Authors: IT Stancil, JE Michalski, D Davis-Hall, HW Chu, JA Park, CM Magin, IV Yang, BJ Smith, E Dobrinskik, DA Schwartz
    Nature Communications, 2021-07-27;12(1):4566.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Long-lasting severe immune dysfunction in Ebola virus disease survivors
    Authors: A Wiedemann, E Foucat, H Hocini, C Lefebvre, BP Hejblum, M Durand, M Krüger, AK Keita, A Ayouba, S Mély, JC Fernandez, A Touré, S Fourati, C Lévy-March, H Raoul, E Delaporte, L Koivogui, R Thiébaut, C Lacabaratz, Y Lévy
    Nat Commun, 2020-07-24;11(1):3730.
    Species: Human
    Sample Types: Serum
  10. AREG mediates the epithelial?mesenchymal transition in pancreatic cancer cells via the EGFR/ERK/NF?&kappaB signalling pathway
    Authors: L Wang, L Wang, H Zhang, J Lu, Z Zhang, H Wu, Z Liang
    Oncol. Rep., 2020-02-27;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. 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, 2019-09-07;0(0):e13027.
    Species: Human
    Sample Types: Serum
  12. 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, 2018-10-15;8(1):14513.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. 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., 2018-02-28;0(0):.
    Species: Human
    Sample Types: Whole Cells
  14. 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, 2017-07-19;12(7):e0181356.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Late acute graft versus host disease: a prospective analysis of clinical outcomes and circulating angiogenic factors
    Blood, 2016-09-13;0(0):.
    Species: Human
    Sample Types: Plasma
  16. 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, 2015-04-27;10(4):e0125793.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. 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, 2015-03-01;35(3):1683-9.
    Species: Human
    Sample Types: Plasma
  18. 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, 2014-03-31;74(10):2869-81.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

  1. Assay Diluent RD1-14 contains visible precipitate. Is this acceptable to use?

    • A small amount of precipitate is expected and acceptable in Assay Diluent RD1-14. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.

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Reviews for Human Amphiregulin Quantikine ELISA Kit

Average Rating: 5 (Based on 6 Reviews)

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Human Amphiregulin Quantikine ELISA Kit
By Anonymous on 11/14/2020
Sample Tested: Prostate cancer

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

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

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

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

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