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

Name: Human Amphiregulin Quantikine ELISA Kit
Brand: R&D Systems
SKU: DAR00
Rating: 5 (6 reviews)

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.

Human Amphiregulin Quantikine ELISA Kit

Catalog #: DAR00
16 Citations 6 Reviews Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
DAR00

Control Products Available

QC40: Quantikine Immunoassay Control Set 1062 Human Amphiregulin

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All Amphiregulin ELISAs

Product Details

Procedure

Citations (16)

FAQs

Supplemental Products

Reviews (6)

Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

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.

Scientific Data

N/A

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Human Amphiregulin ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

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

Related Research Areas

⚠ 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.

Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

50 µL Assay Diluent

Add 50 µL of Assay Diluent to each well.

50 µL Standard, Control, or Sample

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.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

200 µL cold Conjugate

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.
Aspirate and wash 4 times.

200 µL Substrate Solution

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

50 µL Stop Solution

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.

16 Citations: Showing 1 - 10
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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
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
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
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
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
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
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
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
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
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

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

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

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

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

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

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

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.

View all ELISA FAQs