Human 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 human 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 Human 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.
14
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: Mouse
Sample Types: Tissue Homogenates
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The histone demethylase KDM6B fine-tunes the host response to Streptococcus pneumoniae
Authors: MG Connor, TMN Camarasa, E Patey, O Rasid, L Barrio, CM Weight, DP Miller, RS Heyderman, RJ Lamont, J Enninga, MA Hamon
Nature Microbiology, 2020-12-21;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Relative Centrifugal Force (RCF; G-Force) Affects the Distribution of TGF-&beta in PRF Membranes Produced Using Horizontal Centrifugation
Authors: Z Kargarpour, J Nasirzade, L Panahipour, RJ Miron, R Gruber
Int J Mol Sci, 2020-10-15;21(20):.
Species: Human
Sample Types: Cell Culture Supernates
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Induction of interleukin-11 mediated by RhoA GTPase during human cytomegalovirus lytic infection
Authors: S Alarifi, S Alkahtani, AA Al-Qahtani, C Stournaras, G Sourvinos
Cell. Signal., 2020-03-09;70(0):109599.
Species: Human
Sample Types: Cell Culture Supernates
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Thrombin stimulates gene expression and secretion of IL-11 via protease-activated receptor-1 and regulates extravillous trophoblast cell migration
Authors: D Brünnert, I Shekhawat, KR Chahar, J Ehrhardt, J Pandey, JK Yadav, M Zygmunt, P Goyal
J. Reprod. Immunol., 2019-03-09;132(0):35-41.
Species: Human
Sample Types: Cell Culture Supernates
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Human cytomegalovirus replication induces endothelial cell interleukin-11
Authors: KLR Gustafsson, T Renné, C Söderberg-, LM Butler
Cytokine, 2018-05-25;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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New Sequencing technologies help revealing unexpected mutations in Autosomal Dominant Hypercholesterolemia
Authors: S Elbitar, D Susan-Resi, Y Ghaleb, P El Khoury, G Peloso, N Stitziel, JP Rabès, V Carreau, J Hamelin, A Ben-Djoudi, E Bruckert, C Boileau, NG Seidah, M Varret, M Abifadel
Sci Rep, 2018-01-31;8(1):1943.
Species: Human
Sample Types: Cell Lysates
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Fingolimod induces neuroprotective factors in human astrocytes.
Authors: Hoffmann F, Hofereiter J, Rubsamen H, Melms J, Schwarz S, Faber H, Weber P, Putz B, Loleit V, Weber F, Hohlfeld R, Meinl E, Krumbholz M
J Neuroinflammation, 2015-09-30;12(0):184.
Species: Human
Sample Types: Cell Culture Supernates
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Differential reconstitution of T cell subsets following immunodepleting treatment with alemtuzumab (anti-CD52 monoclonal antibody) in patients with relapsing-remitting multiple sclerosis.
Authors: Zhang X, Tao Y, Chopra M, Ahn M, Marcus K, Choudhary N, Zhu H, Markovic-Plese S
J Immunol, 2013-11-06;191(12):5867-74.
Species: Human
Sample Types: Serum
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Chemotherapy stimulates syndecan-1 shedding: a potentially negative effect of treatment that may promote tumor relapse.
Authors: Ramani V, Sanderson R
Matrix Biol, 2013-10-18;35(0):215-22.
Species: Human
Sample Types: Cell Culture Supernates
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Identification of microRNAs inhibiting TGF-beta-induced IL-11 production in bone metastatic breast cancer cells.
Authors: Pollari S, Leivonen SK, Perälä M, Fey V, Kakonen SM, Kallioniemi O
PLoS ONE, 2012-05-21;7(5):e37361.
Species: Human
Sample Types: Cell Culture Supernates
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Interleukin (IL)-4, IL-13, and IL-17A differentially affect the profibrotic and proinflammatory functions of fibrocytes from asthmatic patients.
Authors: Bellini A, Marini MA, Bianchetti L, Barczyk M, Schmidt M, Mattoli S
Mucosal Immunol, 2011-12-21;5(2):140-9.
Species: Human
Sample Types: Cell Culture Supernates
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Heparanase Plays a Dual Role in Driving Hepatocyte Growth Factor (HGF) Signaling by Enhancing HGF Expression and Activity.
Authors: Ramani VC, Yang Y, Ren Y
J. Biol. Chem., 2010-12-03;286(8):6490-9.
Species: Human
Sample Types: Cell Culture Supernates
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Mesenchymal stem cells support migration, extracellular matrix invasion, proliferation, and survival of endothelial cells in vitro.
Authors: Potapova IA, Gaudette GR, Brink PR, Robinson RB, Rosen MR, Cohen IS, Doronin SV
Stem Cells, 2007-03-29;25(7):1761-8.
Species: Human
Sample Types: Cell Culture Supernates
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