Mouse Amphiregulin DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse Amphiregulin. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
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.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Mouse Amphiregulin DuoSet ELISA
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.
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Citations: Showing 1 - 10
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Sustained amphiregulin expression in intermediate alveolar stem cells drives progressive fibrosis
Authors: Zhao, R;Wang, Z;Wang, G;Geng, J;Wu, H;Liu, X;Bin, E;Sui, J;Dai, H;Tang, N;
Cell stem cell
Species: Mouse
Sample Types: Tissue Homogenates, BALF
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The Amphiregulin/EGFR axis has limited contribution in controlling autoimmune diabetes
Authors: Raugh, A;Jing, Y;Bettini, ML;Bettini, M;
Research square
Species: Mouse
Sample Types: Cell Culture Supernates
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Semaphorin 4B is an ADAM17-cleaved adipokine that inhibits adipocyte differentiation and thermogenesis
Authors: A Amin, M Badenes, J Tüshaus, É de Carvalh, E Burbridge, P Faísca, K Trávní?kov, A Barros, S Carobbio, PM Domingos, A Vidal-Puig, LF Moita, S Maguire, K St?íšovský, FJ Ortega, JM Fernández-, SF Lichtentha, C Adrain
Molecular Metabolism, 2023-04-28;0(0):101731.
Species: Mouse
Sample Types: Cell Culture Supernates
Applications: ELISA -
Genome-wide analysis of a cellular exercise model based on electrical pulse stimulation
Authors: B Lee, SK Kim, YJ Shin, YH Son, JW Yang, SM Lee, YR Yang, KP Lee, KS Kwon
Scientific Reports, 2022-12-08;12(1):21251.
Species: Mouse
Sample Types: Cell Culture Supernates
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Macrophage-intrinsic DUOX1 contributes to type 2 inflammation and mucus metaplasia during allergic airway disease
Authors: CR Morris, A Habibovic, CM Dustin, C Schiffers, MC Lin, JL Ather, YMW Janssen-He, ME Poynter, O Utermohlen, M Krönke, A van der Vl
Mucosal Immunology, 2022-06-02;0(0):.
Species: Mouse
Sample Types: BALF
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Antigen receptor-engineered Tregs inhibit CNS autoimmunity in cell therapy using non-redundant immune mechanisms in mice
Authors: J Pohar, R O'Connor, B Manfroi, ME Behi, L Jouneau, P Boudinot, M Bunse, W Uckert, M Luka, M Ménager, R Liblau, SM Anderton, S Fillatreau
European Journal of Immunology, 2022-05-27;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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A high docosahexaenoic acid diet alters lung inflammation and recovery following repetitive exposure to aqueous organic dust extracts: Role of DHA in repetitive organic dust exposure
Authors: A Ulu, A Burr, AJ Heires, J Pavlik, T Larsen, PA Perez, C Bravo, NV DiPatrizio, M Baack, DJ Romberger, TM Nordgren
The Journal of nutritional biochemistry, 2021-06-12;0(0):108797.
Species: Mouse
Sample Types: BALF
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In vivo silencing of amphiregulin by a novel effective Self-Assembled-Micelle inhibitory RNA ameliorates renal fibrosis via inhibition of EGFR signals
Authors: SS Son, S Hwang, JH Park, Y Ko, SI Yun, JH Lee, B Son, TR Kim, HO Park, EY Lee
Scientific Reports, 2021-01-26;11(1):2191.
Species: Mouse
Sample Types: Tissue Homogenates
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Genes adapt to outsmart gene-targeting strategies in mutant mouse strains by skipping exons to reinitiate transcription and translation
Authors: V Hosur, BE Low, D Li, GA Stafford, V Kohar, LD Shultz, MV Wiles
Genome Biol., 2020-07-09;21(1):168.
Species: Mouse
Sample Types: Cell Culture Supernates
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MyD88 regulates a prolonged adaptation response to environmental dust exposure-induced lung disease
Authors: AN Johnson, JR Harkema, AJ Nelson, JD Dickinson, J Kalil, MJ Duryee, GM Thiele, B Kumar, AB Singh, R Gaurav, SC Glover, Y Tang, DJ Romberger, T Kielian, JA Poole
Respir. Res., 2020-04-22;21(1):97.
Species: Mouse
Sample Types: BALF
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AhR controls redox homeostasis and shapes the tumor microenvironment in BRCA1-associated breast cancer
Authors: SP Kubli, C Bassi, C Roux, A Wakeham, C Göbl, W Zhou, SM Jafari, B Snow, L Jones, L Palomero, KL Thu, L Cassetta, D Soong, T Berger, P Ramachandr, SP Baniasadi, G Duncan, M Lindzen, Y Yarden, C Herranz, C Lazaro, MF Chu, J Haight, P Tinto, J Silvester, DW Cescon, A Petit, S Pettersson, JW Pollard, TW Mak, MA Pujana, P Cappello, C Gorrini
Proc. Natl. Acad. Sci. U.S.A., 2019-02-07;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Deoxycholic acid activates epidermal growth factor receptor and promotes intestinal carcinogenesis by ADAM17-dependent ligand release
Authors: W Dong, L Liu, Y Dou, M Xu, T Liu, S Wang, Y Zhang, B Deng, B Wang, H Cao
J. Cell. Mol. Med., 2018-06-29;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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ADAM17 is required for EGF-R-induced intestinal tumors via IL-6 trans-signaling
Authors: S Schmidt, N Schumacher, J Schwarz, S Tangermann, L Kenner, M Schlederer, M Sibilia, M Linder, A Altendorf-, T Knösel, ES Gruber, G Oberhuber, J Bolik, A Rehman, A Sinha, J Lokau, P Arnold, AS Cabron, F Zunke, C Becker-Pau, A Preaudet, P Nguyen, J Huynh, S Afshar-Ste, AL Chand, J Westermann, PJ Dempsey, C Garbers, D Schmidt-Ar, P Rosenstiel, T Putoczki, M Ernst, S Rose-John
J. Exp. Med., 2018-02-22;0(0):.
Species: Mouse
Sample Types: Tissue Culture Supernates
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Genetic deletion of amphiregulin restores the normal skin phenotype in a mouse model of the human skin disease tylosis
Authors: V Hosur, BE Low, LD Shultz, MV Wiles
Biol Open, 2017-08-15;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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DUOX1 mediates persistent epithelial EGFR activation, mucous cell metaplasia, and airway remodeling during allergic asthma
JCI Insight, 2016-11-03;1(18):e88811.
Species: Mouse
Sample Types: BALF
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Progesterone-Based Therapy Protects Against Influenza by Promoting Lung Repair and Recovery in Females
PLoS Pathog, 2016-09-15;12(9):e1005840.
Species: Mouse
Sample Types: Tissue Homogenates
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ADAM17 substrate release in proximal tubule drives kidney fibrosis
JCI Insight, 2016-08-18;1(13):.
Species: Mouse
Sample Types: Tissue Homogenates
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RAF inhibitors that evade paradoxical MAPK pathway activation.
Authors: Zhang C, Spevak W, Zhang Y, Burton E, Ma Y, Habets G, Zhang J, Lin J, Ewing T, Matusow B, Tsang G, Marimuthu A, Cho H, Wu G, Wang W, Fong D, Nguyen H, Shi S, Womack P, Nespi M, Shellooe R, Carias H, Powell B, Light E, Sanftner L, Walters J, Tsai J, West B, Visor G, Rezaei H, Lin P, Nolop K, Ibrahim P, Hirth P, Bollag G
Nature, 2015-10-14;526(7574):583-6.
Species: Mouse
Sample Types: Cell Culture Supernates
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Delivery of GM-CSF to Protect against Influenza Pneumonia.
Authors: Subramaniam R, Hillberry Z, Chen H, Feng Y, Fletcher K, Neuenschwander P, Shams H
PLoS ONE, 2015-04-29;10(4):e0124593.
Species: Mouse
Sample Types: BALF
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Asthma increases susceptibility to heterologous but not homologous secondary influenza.
Authors: Furuya Y, Roberts S, Hurteau G, Sanfilippo A, Racine R, Metzger D
J Virol, 2014-06-04;88(16):9166-81.
Species: Mouse
Sample Types: BALF
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IL-9-mediated survival of type 2 innate lymphoid cells promotes damage control in helminth-induced lung inflammation.
Authors: Turner, Jan-Eric, Morrison, Peter J, Wilhelm, Christop, Wilson, Mark, Ahlfors, Helena, Renauld, Jean-Chr, Panzer, Ulf, Helmby, Helena, Stockinger, Brigitta
J Exp Med, 2013-11-18;210(13):2951-65.
Species: Mouse
Sample Types: Cell Culture Supernates
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Limited anti-inflammatory role for interleukin-1 receptor like 1 (ST2) in the host response to murine postinfluenza pneumococcal pneumonia.
Authors: Blok D, van der Sluijs K, Florquin S, de Boer O, van 't Veer C, de Vos A, van der Poll T
PLoS ONE, 2013-03-06;8(3):e58191.
Species: Mouse
Sample Types: Tissue Homogenates
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Amphiregulin, an epidermal growth factor receptor ligand, plays an essential role in the pathogenesis of transforming growth factor-beta-induced pulmonary fibrosis.
Authors: Zhou Y, Lee J, Lee C, Cho W, Kang M, Koff J, Yoon P, Chae J, Park H, Elias J, Lee C
J Biol Chem, 2012-10-19;287(50):41991-2000.
Species: Mouse
Sample Types: BALF
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Innate lymphoid cells promote lung-tissue homeostasis after infection with influenza virus.
Authors: Monticelli LA, Sonnenberg GF, Abt MC
Nat. Immunol., 2011-11-01;12(11):1045-54.
Species: Mouse
Sample Types: Cell Culture Supernates
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Ectodomain shedding of EGFR ligands and TNFR1 dictates hepatocyte apoptosis during fulminant hepatitis in mice.
Authors: Murthy A, Defamie V, Smookler DS
J. Clin. Invest., 2010-07-12;120(8):2731-44.
Species: Mouse
Sample Types: Serum
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