Mouse IL-1 alpha/IL-1F1 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-1alpha/IL-1F1. 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-1 alpha/IL-1F1
IL-1 alpha (Interleukin-1 alpha) is a cytokine that is induced during microbial exposure and promotes the release of multiple inflammatory mediators and acute phase proteins. It also supports hematopoietic stem cell expansion, promotes neutrophil mobilization from the bone marrow, and promotes osteoclast formation. IL-1 alpha signals through a receptor complex composed of IL-1 RI and IL-1 RAcP. It also binds the decoy receptor IL-1 RII which associates with IL-1 RAcP but does not transduce 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-1 alpha/IL-1F1 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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Microglial NLRP3 inflammasome activates neurotoxic astrocytes in depression-like mice
Authors: S Li, Y Fang, Y Zhang, M Song, X Zhang, X Ding, H Yao, M Chen, Y Sun, J Ding, Q Wang, M Lu, G Wu, G Hu
Cell Reports, 2022-10-25;41(4):111532.
Species: Mouse
Sample Types: Cell Culture Supernates
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Smoking status impacts treatment efficacy in smoke-induced lung inflammation: A pre-clinical study
Authors: N Milad, M Pineault, F Tremblay, J Routhier, A Lechasseur, MJ Beaulieu, S Aubin, MC Morissette
Frontiers in Pharmacology, 2022-09-07;13(0):971238.
Species: Mouse
Sample Types: BALF
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Acinetobacter quorum sensing contributes to inflammation-induced inhibition of orthopaedic implant osseointegration
Authors: H Choe, BS Hausman, KM Hujer, O Akkus, PN Rather, Z Lee, RA Bonomo, EM Greenfield
Oncogene, 2022-06-09;43(0):267-276.
Species: Mouse
Sample Types: Tissue Homogenates
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Heterocellular OSM-OSMR signalling reprograms fibroblasts to promote pancreatic cancer growth and metastasis
Authors: BY Lee, EKJ Hogg, CR Below, A Kononov, A Blanco-Gom, F Heider, J Xu, C Hutton, X Zhang, T Scheidt, K Beattie, A Lamarca, M McNamara, JW Valle, C Jørgensen
Nature Communications, 2021-12-17;12(1):7336.
Species: Mouse
Sample Types: Tissue Homogenates
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NLRP1 acts as a negative regulator of Th17 cell programming in mice and humans with autoimmune diabetes
Authors: FRC Costa, JA Leite, DM Rassi, JF da Silva, J Elias-Oliv, JB Guimarães, MC Foss-Freit, NOS Câmara, A Pontillo, RC Tostes, JS Silva, D Carlos
Cell Reports, 2021-05-25;35(8):109176.
Species: Mouse
Sample Types: Tissue Homogenates
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&beta-Glucan-stimulated neutrophil secretion of IL-1&alpha is independent of GSDMD and mediated through extracellular vesicles
Authors: B Ratitong, M Marshall, E Pearlman
Cell Reports, 2021-05-18;35(7):109139.
Species: Mouse, Transgenic Mouse
Sample Types: Cell Culture Supernates
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A NOVEL NOX/PHOX-CD38-NAADP-TFEB AXIS IMPORTANT FOR MACROPHAGE ACTIVATION DURING BACTERIAL PHAGOCYTOSIS
Authors: M Najibi, HH Honwad, JA Moreau, SM Becker, JE Irazoqui
Autophagy, 2021-04-13;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Tim4 recognizes carbon nanotubes and mediates phagocytosis leading to granuloma formation
Authors: S Omori, M Tsugita, Y Hoshikawa, M Morita, F Ito, SI Yamaguchi, Q Xie, O Noyori, T Yamaguchi, A Takada, T Saitoh, S Toyokuni, H Akiba, S Nagata, K Kinoshita, M Nakayama
Cell Reports, 2021-02-09;34(6):108734.
Species: Mouse
Sample Types: Cell Culture Supernates
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Gasdermin D restricts Burkholderia cenocepacia infection in vitro and in vivo
Authors: S Estfanous, K Krause, MNK Anne, M Eltobgy, K Caution, A Abu Khweek, K Hamilton, A Badr, K Daily, C Carafice, D Baetzhold, X Zhang, T Li, H Wen, MA Gavrilin, H Haffez, S Soror, AO Amer
Scientific Reports, 2021-01-13;11(1):855.
Species: Mouse
Sample Types: Cell Culture Supernates
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The Pseudomonas aeruginosa protease LasB directly activates IL-1&beta
Authors: J Sun, DL LaRock, EA Skowronski, JM Kimmey, J Olson, Z Jiang, AJ O'Donoghue, V Nizet, CN LaRock
EBioMedicine, 2020-09-23;60(0):102984.
Species: Mouse
Sample Types: Cell Culture Supernates
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Interleukin-1 Receptor-Induced Nitric Oxide Production in the Pancreas Controls Hyperglycemia Caused by Scorpion Envenomation
Authors: MB Reis, J Elias-Oliv, MR Pastore, SG Ramos, LG Gardinassi, LH Faccioli
Toxins (Basel), 2020-03-05;12(3):.
Species: Mouse
Sample Types: Serum
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The three cytokines IL-1?, IL-18, and IL-1? share related but distinct secretory routes
Authors: VS Tapia, MJD Daniels, P Palazón-Ri, M Dewhurst, NM Luheshi, J Rivers-Aut, J Green, E Redondo-Ca, P Kaldis, G Lopez-Cast, D Brough
J. Biol. Chem., 2019-04-02;294(21):8325-8335.
Species: Mouse
Sample Types: Cell Culture Supernates
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Mycobacterium tuberculosis carrying a rifampicin drug resistance mutation reprograms macrophage metabolism through cell wall lipid changes
Authors: NC Howard, ND Marin, M Ahmed, BA Rosa, J Martin, M Bambouskov, A Sergushich, E Loginichev, N Kurepina, J Rangel-Mor, L Chen, BN Kreiswirth, RS Klein, JM Balada-Lla, JB Torrelles, GK Amarasingh, M Mitreva, MN Artyomov, FF Hsu, B Mathema, SA Khader
Nat Microbiol, 2018-09-17;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Activation of hypoxia-inducible factor�1 attenuates periapical inflammation and bone loss
Authors: K Hirai, H Furusho, K Hirota, H Sasaki
Int J Oral Sci, 2018-04-13;10(2):12.
Species: Mouse
Sample Types: Cell Culture Supernates
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Saturated Fatty Acids Undergo Intracellular Crystallization and Activate the NLRP3 Inflammasome in Macrophages
Authors: T Karasawa, A Kawashima, F Usui-Kawan, S Watanabe, H Kimura, R Kamata, K Shirasuna, Y Koyama, A Sato-Tomit, T Matsuzaka, H Tomoda, SY Park, N Shibayama, H Shimano, T Kasahara, M Takahashi
Arterioscler. Thromb. Vasc. Biol., 2018-02-08;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Human caspase-4 detects tetra-acylated LPS and cytosolic Francisella and functions differently from murine caspase-11
Authors: B Lagrange, S Benaoudia, P Wallet, F Magnotti, A Provost, F Michal, A Martin, F Di Lorenzo, BF Py, A Molinaro, T Henry
Nat Commun, 2018-01-16;9(1):242.
Species: Mouse
Sample Types: Cell Culture Supernates
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CRISPR/Cas9 mediated mutation of mouse IL-1? nuclear localisation sequence abolishes expression
Authors: MJD Daniels, AD Adamson, N Humphreys, D Brough
Sci Rep, 2017-12-06;7(1):17077.
Species: Mouse
Sample Types: Cell Culture Supernates
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Caspase-2 deficiency accelerates chemically induced liver cancer in mice
Cell Death Differ, 2016-08-12;0(0):.
Species: Mouse
Sample Types: Serum
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Reducing hypoxia and inflammation during invasive pulmonary aspergillosis by targeting the Interleukin-1 receptor
Sci Rep, 2016-05-24;6(0):26490.
Species: Mouse
Sample Types: Tissue Homogenates
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Infection-Mediated Priming of Phagocytes Protects against Lethal Secondary Aspergillus fumigatus Challenge
Authors: A Savers, O Rasid, M Parlato, M Brock, G Jouvion, B Ryffel, JM Cavaillon, G Eberl, O Ibrahim-Gr
PLoS ONE, 2016-04-14;11(4):e0153829.
Species: Mouse
Sample Types: Tissue Culture Supernates
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The Inflammatory Caspases-1 and -11 Mediate the Pathogenesis of Dermatitis in Sharpin-Deficient Mice.
Authors: Douglas T, Champagne C, Morizot A, Lapointe J, Saleh M
J Immunol, 2015-07-27;195(5):2365-73.
Species: Mouse
Sample Types: Cell Culture Supernates
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A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases.
Authors: Coll R, Robertson A, Chae J, Higgins S, Munoz-Planillo R, Inserra M, Vetter I, Dungan L, Monks B, Stutz A, Croker D, Butler M, Haneklaus M, Sutton C, Nunez G, Latz E, Kastner D, Mills K, Masters S, Schroder K, Cooper M, O'Neill L
Nat Med, 2015-02-16;21(3):248-55.
Species: Mouse
Sample Types: Cell Culture Supernates
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Dendritic cell IL-1alpha and IL-1beta are polyubiquitinated and degraded by the proteasome.
Authors: Ainscough J, Frank Gerberick G, Zahedi-Nejad M, Lopez-Castejon G, Brough D, Kimber I, Dearman R
J Biol Chem, 2014-11-04;289(51):35582-92.
Species: Mouse
Sample Types: Cell Lysates
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A conservative amino acid mutation in the master regulator FleQ renders Pseudomonas aeruginosa aflagellate.
Authors: Jain, Ruchi, Kazmierczak, Barbara
PLoS ONE, 2014-05-14;9(5):e97439.
Species: Mouse
Sample Types: BALF
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Animal model of respiratory syncytial virus: CD8+ T cells cause a cytokine storm that is chemically tractable by sphingosine-1-phosphate 1 receptor agonist therapy.
Authors: Walsh K, Teijaro J, Brock L, Fremgen D, Collins P, Rosen H, Oldstone M
J Virol, 2014-03-26;88(11):6281-93.
Species: Mouse
Sample Types: BALF
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Staphylococcus aureus infection of mice expands a population of memory gammadelta T cells that are protective against subsequent infection.
Authors: Murphy A, O'Keeffe K, Lalor S, Maher B, Mills K, McLoughlin R
J Immunol, 2014-03-12;192(8):3697-708.
Species: Mouse
Sample Types: Peritoneal Lavage Fluid
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Signaling via tumor necrosis factor receptor 1 but not Toll-like receptor 2 contributes significantly to hydrosalpinx development following Chlamydia muridarum infection.
Authors: Dong X, Liu Y, Chang X, Lei L, Zhong G
Infect Immun, 2014-02-18;82(5):1833-9.
Species: Mouse
Sample Types: Cell Culture Supernates
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MyD88 signaling regulates both host defense and immunopathogenesis during pneumocystis infection.
Authors: Bello-Irizarry S, Wang J, Johnston C, Gigliotti F, Wright T
J Immunol, 2013-11-29;192(1):282-92.
Species: Mouse
Sample Types: BALF
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Mesenchymal stem cells inhibit cutaneous radiation-induced fibrosis by suppressing chronic inflammation.
Authors: Horton J, Hudak K, Chung E, White A, Scroggins B, Burkeen J, Citrin D
Stem Cells, 2013-10-01;31(10):2231-41.
Species: Mouse
Sample Types: Tissue Homogenates
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Endothelial cells are central orchestrators of cytokine amplification during influenza virus infection.
Authors: Teijaro JR, Walsh KB, Cahalan S, Fremgen DM, Roberts E, Scott F, Martinborough E, Peach R, Oldstone MB, Rosen H
Cell, 2011-09-16;146(6):980-91.
Species: Mouse
Sample Types: BALF
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CD4 T cells promote rather than control tuberculosis in the absence of PD-1-mediated inhibition.
Authors: Barber DL, Mayer-Barber KD, Feng CG
J. Immunol., 2010-12-20;186(3):1598-607.
Species: Mouse
Sample Types: BALF
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MyD88 regulation of Fusarium keratitis is dependent on TLR4 and IL-1R1 but not TLR2.
Authors: Tarabishy AB, Aldabagh B, Sun Y, Imamura Y, Mukherjee PK, Lass JH, Ghannoum MA, Pearlman E
J. Immunol., 2008-07-01;181(1):593-600.
Species: Mouse
Sample Types: Tissue Homogenates
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Lack of Toll IL-1R8 exacerbates Th17 cell responses in fungal infection.
Authors: Bozza S, Zelante T, Moretti S, Bonifazi P, DeLuca A, D'Angelo C, Giovannini G, Garlanda C, Boon L, Bistoni F, Puccetti P, Mantovani A, Romani L
J. Immunol., 2008-03-15;180(6):4022-31.
Species: Mouse
Sample Types: Cell Culture Supernates
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Caspase-1 contributes to Chlamydia trachomatis-induced upper urogenital tract inflammatory pathologies without affecting the course of infection.
Authors: Cheng W, Shivshankar P, Li Z, Chen L, Yeh IT, Zhong G
Infect. Immun., 2007-11-19;76(2):515-22.
Species: Mouse
Sample Types: Cell Culture Supernates
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Interleukin 10 regulates inflammatory cytokine synthesis to protect against lipopolysaccharide-induced abortion and fetal growth restriction in mice.
Authors: Robertson SA, Care AS, Skinner RJ
Biol. Reprod., 2007-01-10;76(5):738-48.
Species: Mouse
Sample Types: Serum
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Essential role for IL-10 in resistance to lipopolysaccharide-induced preterm labor in mice.
Authors: Robertson SA, Skinner RJ, Care AS
J. Immunol., 2006-10-01;177(7):4888-96.
Species: Mouse
Sample Types: Serum
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