Mouse Amphiregulin DuoSet ELISA

Catalog # Availability Size / Price Qty
DY989
Ancillary Products Available
Amphiregulin ELISA Kit ELISA
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Product Details
Procedure
Citations (13)
FAQs
Supplemental Products
Reviews (3)

Mouse Amphiregulin DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* 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)

Data Example

Mouse Amphiregulin ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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.

13 Citations: Showing 1 - 10
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  1. 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;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. 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;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  3. DUOX1 mediates persistent epithelial EGFR activation, mucous cell metaplasia, and airway remodeling during allergic asthma
    JCI Insight, 2016;1(18):e88811.
    Species: Mouse
    Sample Types: BALF
  4. ADAM17 substrate release in proximal tubule drives kidney fibrosis
    JCI Insight, 2016;1(13):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  5. Progesterone-Based Therapy Protects Against Influenza by Promoting Lung Repair and Recovery in Females
    PLoS Pathog, 2016;12(9):e1005840.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. 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;526(7574):583-6.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. 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;10(4):e0124593.
    Species: Mouse
    Sample Types: BALF
  8. 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;88(16):9166-81.
    Species: Mouse
    Sample Types: BALF
  9. 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;210(13):2951-65.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. 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;8(3):e58191.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. 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;287(50):41991-2000.
    Species: Mouse
    Sample Types: BALF
  12. Innate lymphoid cells promote lung-tissue homeostasis after infection with influenza virus.
    Authors: Monticelli LA, Sonnenberg GF, Abt MC
    Nat. Immunol., 2011;12(11):1045-54.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. 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;120(8):2731-44.
    Species: Mouse
    Sample Types: Serum

FAQs

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Reviews for Mouse Amphiregulin DuoSet ELISA

Average Rating: 5 (Based on 3 Reviews)

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Mouse Amphiregulin DuoSet ELISA
By Anonymous on 04/18/2018
Sample Tested: LL/2 mouse Lewis lung carcinoma cell line

Mouse Amphiregulin DuoSet ELISA
By Anonymous on 04/05/2018
Sample Tested: mouse pancreas tissue

Mouse Amphiregulin DuoSet ELISA
By Anonymous on 04/02/2018
Sample Tested: mouse pancreatic cancer tissue,human pancreatic cancer