Mouse IL-11 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 IL-11. 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: IL-11
Interleukin-11, also known as AGIF, is a pleiotropic cytokine in the IL-6 family, which also includes LIF, CNTF, Oncostatin M, Cardiotrophin-1, IL-27, and IL-31. It is secreted by osteoblasts, synoviocytes, fibroblasts, chondrocytes, intestinal myofibroblasts, and trophoblasts. It is found in the plasma mainly during inflammation, and is considered to be primarily anti-inflammatory. It stimulates hematopoiesis and thrombopoiesis, regulates macrophage differentiation, and confers mucosal protection in the intestine. It has also been found to enhance T cell polarization toward Th2, promote B cell IgG production, increase osteoclast bone absorption, protect endothelial cells from oxidative stress, and regulate epithelial proliferation and apoptosis. IL-11 signals through a receptor complex that contains IL-11 R alpha and gp130. A soluble form of IL-11 R alpha can bind IL-11 and either form a signaling complex with gp130 on the cell surface or inhibit cell surface IL-11 R alpha/gp130 signaling.
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 IL-11 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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Interleukin-11 Suppresses Ocular Surface Inflammation and Accelerates Wound Healing
Authors: Singh, A;Cho, WJ;Pulimamidi, VK;Mittal, SK;Chauhan, SK;
Investigative ophthalmology & visual science
Species: Human
Sample Types: Cell Culture Supernates
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Hepatocyte Specific gp130 Signalling Underlies APAP Induced Liver Injury
Authors: J Dong, WW Lim, SG Shekeran, J Tan, SY Lim, JWT Goh, BL George, S Schafer, SA Cook, AA Widjaja
International Journal of Molecular Sciences, 2022-06-25;23(13):.
Species: Mouse
Sample Types: Serum
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Hepatocyte-specific IL11 cis-signaling drives lipotoxicity and underlies the transition from NAFLD to NASH
Authors: J Dong, S Viswanatha, E Adami, BK Singh, SP Chothani, B Ng, WW Lim, J Zhou, M Tripathi, NSJ Ko, SG Shekeran, J Tan, SY Lim, M Wang, PM Lio, PM Yen, S Schafer, SA Cook, AA Widjaja
Nature Communications, 2021-01-04;12(1):66.
Species: Mouse
Sample Types: Serum
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Prolonged infection triggered by dormant Mycobacterium tuberculosis: Immune and inflammatory responses in lungs of genetically susceptible and resistant mice
Authors: T Kondratiev, M Shleeva, M Kapina, E Rubakova, I Linge, A Dyatlov, E Kondratiev, A Kaprelyant, A Apt
PLoS ONE, 2020-09-24;15(9):e0239668.
Species: Mouse
Sample Types: Cell Culture Supernates
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Endoplasmic reticulum stress differentially modulates the IL-6 family of cytokines in murine astrocytes and macrophages
Authors: CL Sanchez, SG Sims, JD Nowery, GP Meares
Sci Rep, 2019-10-17;9(1):14931.
Species: Mouse
Sample Types: Cell Culture Supernates
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IL-11 Induces Encephalitogenic Th17 Cells in Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis
Authors: X Zhang, N Kiapour, S Kapoor, T Khan, M Thamilaras, Y Tao, S Cohen, R Miller, RA Sobel, S Markovic-P
J. Immunol., 2019-07-24;0(0):.
Species: Mouse
Sample Types: Serum
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Neuroprotective effects of ellagic acid on cuprizone-induced acute demyelination through limitation of microgliosis, adjustment of CXCL12/IL-17/IL-11 axis and restriction of mature oligodendrocytes apoptosis
Authors: N Sanadgol, F Golab, Z Tashakkor, N Taki, S Moradi Kou, A Mostafaie, M Mehdizadeh, M Abdollahi, G Taghizadeh, M Sharifzade
Pharm Biol, 2017-12-01;55(1):1679-1687.
Species: Mouse
Sample Types: Tissue Homogenates
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Critical contribution of nuclear factor erythroid 2-related factor 2 (NRF2) to electrophile-induced Interleukin-11 production
Authors: Hiroyasu Nakano
J. Biol. Chem., 2016-11-21;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Optimal attenuation of experimental autoimmune encephalomyelitis by intravenous immunoglobulin requires an intact interleukin-11 receptor.
Authors: Figueiredo C, Drohomyrecky P, McCarthy S, Leontyev D, Ma X, Branch D, Dunn S
PLoS ONE, 2014-07-31;9(7):e101947.
Species: Mouse
Sample Types: Serum
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Evaluating the role of IL-11, a novel cytokine in the IL-6 family, in a mouse model of spinal cord injury.
Authors: Cho N, Nguyen DH, Satkunendrarajah K, Branch DR, Fehlings MG
J Neuroinflammation, 2012-06-20;9(1):134.
Species: Mouse
Sample Types: Tissue Homogenates
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Critical role of the disintegrin metalloprotease ADAM17 for intestinal inflammation and regeneration in mice.
Authors: Chalaris A, Adam N, Sina C, Rosenstiel P, Lehmann-Koch J, Schirmacher P, Hartmann D, Cichy J, Gavrilova O, Schreiber S, Jostock T, Matthews V, Hasler R, Becker C, Neurath MF, Reiss K, Saftig P, Scheller J, Rose-John S
J. Exp. Med., 2010-07-05;207(8):1617-24.
Species: Mouse
Sample Types: Cell Culture Supernates
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