Human 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 human 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 Human 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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Modulating NLRP3 splicing with antisense oligonucleotides to control pathological inflammation
Authors: Klein, R;Onyuru, J;Viera, EM;Putnam, CD;Hoffman, HM;Hastings, ML;
bioRxiv : the preprint server for biology
Species: Human
Sample Types: Cell Culture Supernates
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Effect of progestin-based contraceptives on HIV-associated vaginal immune biomarkers and microbiome in adolescent girls
Authors: Nasr, MA;Aldous, A;Daniels, J;Joy, C;Capozzi, E;Yang, M;Moriarty, P;Emmanuel-Baker, V;Malcolm, S;Green, SJ;Gomez-Lobo, V;Ghosh, M;
PloS one
Species: Human
Sample Types: Vaginal Swab
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Transcriptomic changes during the replicative senescence of human articular chondrocytes
Authors: Atasoy-Zeybek, A;Hawse, GP;Nagelli, CV;De Padilla, CL;Abdel, MP;Evans, CH;
bioRxiv : the preprint server for biology
Species: Human
Sample Types: Cell Culture Supernates
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Ventilatory support and inflammatory peptides in hospitalised patients with COVID-19: A prospective cohort trial
Authors: Gysan, MR;Milacek, C;Bal, C;Zech, A;Brugger, J;Milos, RI;Antoniewicz, L;Idzko, M;Gompelmann, D;
PloS one
Species: Human
Sample Types: Serum
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Serum interleukin-6, procalcitonin, and C-reactive protein at hospital admission can identify patients at low risk for severe COVID-19 progression
Authors: Zobel, CM;Wenzel, W;Krüger, JP;Baumgarten, U;Wagelöhner, T;Neumann, N;Foroutan, B;Müller, R;Müller, A;Rauschning, D;Schü beta ler, M;Scheit, L;Weinreich, F;Oltmanns, K;Keidel, F;Koch, M;Spethmann, S;Schreiner, M;
Frontiers in microbiology
Species: Human
Sample Types: Serum
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Human TH17 cells engage gasdermin E pores to release IL-1alpha on NLRP3 inflammasome activation
Authors: YY Chao, A Puhach, D Frieser, M Arunkumar, L Lehner, T Seeholzer, A Garcia-Lop, M van der Wa, S Fibi-Smeta, A Dietschman, T Sommermann, T ?ikovi?, L Taher, MS Gresnigt, SJ Vastert, F van Wijk, G Panagiotou, D Krappmann, O Gro beta, CE Zielinski
Nature Immunology, 2023-01-05;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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A whole genome CRISPR screen identifies AHR loss as a mechanism of PARP7 inhibitor resistance
Authors: H Chen, ME Diolaiti, PC O'Leary, A Rojc, NJ Krogan, M Kim, A Ashworth
Molecular Cancer Therapeutics, 2022-07-05;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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A prospective cohort study providing insights for markers of adverse pregnancy outcome in older mothers
Authors: SC Lean, RL Jones, SA Roberts, AEP Heazell
BMC pregnancy and childbirth, 2021-10-20;21(1):706.
Species: Human
Sample Types: Serum
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Skin Sensitization Evaluation of Carbon-Based Graphene Nanoplatelets
Authors: SH Kim, SH Hong, JH Lee, DH Lee, K Jung, JY Yang, HS Shin, J Lee, J Jeong, JH Oh
Toxics, 2021-03-17;9(3):.
Species: Human
Sample Types: cell culture supernatant
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Synthesis of Retinol-Loaded Lipid Nanocarrier via Vacuum Emulsification to Improve Topical Skin Delivery
Authors: SH Jun, H Kim, H Lee, JE Song, SG Park, NG Kang
Polymers, 2021-03-08;13(5):.
Species: Human
Sample Types: Cell Culture Supernates
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Interleukin-1&alpha Is a Critical Mediator of the Response of Human Bronchial Fibroblasts to Eosinophilic Inflammation
Authors: K Bernau, JP Leet, H Floerke, EM Bruhn, AL Noll, IS McDermott, S Esnault, NN Jarjour, N Sandbo
Cells, 2021-03-02;10(3):.
Species: Human
Sample Types: BALF
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Acid-electrolyzed functional water-induces Interleukin-1&alpha release from Intracellular Storage Sites in Oral Squamous Cell Carcinoma
Authors: T Takemoto, R Kaetsu, M Hanayama, Y Ishiyama, M Sadamura, K Nishio, M Tsunoda, M Asano, M Motoyoshi
International Journal of Medical Sciences, 2021-02-18;18(8):1746-1752.
Species: Human
Sample Types: Cell Culture Supernates
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ROCK inhibition modulates the senescence-associated secretory phenotype (SASP) in oral keratinocytes
Authors: S Niklander, D Bandaru, DW Lambert, KD Hunter
FEBS Open Bio, 2020-11-06;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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The Delivery of &alpha1-Antitrypsin Therapy Through Transepidermal Route: Worthwhile to Explore
Authors: S Tumpara, B Martinez-D, G Gomez-Mari, B Liu, DS DeLuca, E Korenbaum, D Jonigk, F Jugert, FM Wurm, MJ Wurm, T Welte, S Janciauski
Front Pharmacol, 2020-07-03;11(0):983.
Species: Human
Sample Types: Cell Culture Supernates
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Cytotoxic and Anti-Inflammatory Effects of Ent-Kaurane Derivatives Isolated from the Alpine Plant Sideritis hyssopifolia
Authors: A Aimond, K Calabro, C Audoin, E Olivier, M Dutot, P Buron, P Rat, O Laprévote, S Prado, E Roulland, OP Thomas, G Genta-Jouv
Molecules, 2020-01-29;25(3):.
Species: Human
Sample Types: Cell Culture Supernates
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Ouabain signaling in oral squamous cell carcinoma cells
Authors: S Saito, M Ohtsu, M Asano, T Ishigami
J Oral Sci, 2019-09-20;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Inflammasome-Driven Interleukin-1α and�Interleukin-1? Production in Atherosclerotic Plaques Relates to Hyperlipidemia and Plaque Complexity
Authors: X Jiang, F Wang, Y Wang, A Gisterå, J Roy, G Paulsson-B, U Hedin, A Lerman, GK Hansson, J Herrmann, ZQ Yan
JACC Basic Transl Sci, 2019-06-24;4(3):304-317.
Species: Human
Sample Types: Tissue Culture Supernates
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Cytokine Diversity in Human Peripheral Blood Eosinophils: Profound Variability of IL-16
Authors: M Ma, CM Percopo, DE Sturdevant, AC Sek, HD Komarow, HF Rosenberg
J. Immunol., 2019-06-10;0(0):.
Species: Human
Sample Types: Cell Lysates
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Newly defined ABCB5+ dermal mesenchymal stem cells promote healing of chronic iron overload wounds via secretion of interleukin-1 receptor antagonist
Authors: S Vander Bek, JC de Vries, B Meier-Schi, P Meyer, D Jiang, A Sindrilaru, FF Ferreira, A Hainzl, S Schatz, J Muschhamme, NJ Scheurmann, P Kampilafko, AM Seitz, L Dürselen, A Ignatius, MA Kluth, C Ganss, M Wlaschek, K Singh, P Maity, NY Frank, MH Frank, K Scharffett
Stem Cells, 2019-05-13;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Regulation of radiosensitivity by 4-methylumbelliferone via the suppression of interleukin-1 in fibrosarcoma cells
Authors: R Saga, K Hasegawa, K Murata, M Chiba, T Nakamura, K Okumura, E Tsuruga, Y Hosokawa
Oncol Lett, 2019-01-29;17(3):3555-3561.
Species: Human
Sample Types: Cell Culture Supernates
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A Transient Pseudosenescent Secretome Promotes Tumor Growth after Antiangiogenic Therapy Withdrawal
Authors: M Mastri, A Tracz, CR Lee, M Dolan, K Attwood, JG Christense, S Liu, JML Ebos
Cell Rep, 2018-12-26;25(13):3706-3720.e8.
Species: Human
Sample Types: Cell Culture Supernates
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Modulation of salivary cytokines in response to alcohol, tobacco and caffeine consumption: a pilot study
Authors: CC Sheth, RM López-Pedr, MDM Jovani-San, R González-M, V Veses
Sci Rep, 2018-11-12;8(1):16687.
Species: Human
Sample Types: Saliva
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Acid-electrolyzed functional water induces extracellular matrix metalloproteinase inducer, a possible novel alarmin, secretion from oral squamous cell carcinoma cell lines
Authors: M Kusunoki, E Sata, K Nishio, T Tanaka, T Nishida, N Sugano, S Sato, M Asano
Int J Med Sci, 2018-09-07;15(12):1365-1372.
Species: Human
Sample Types: Cell Culture Supernates
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The Effects of Varying Degree of MWCNT Carboxylation on Bioactivity in Various In Vivo and In Vitro Exposure Models
Authors: RF Hamilton, Z Wu, S Mitra, A Holian
Int J Mol Sci, 2018-01-25;19(2):.
Species: Human
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: Human
Sample Types: Cell Culture Supernates
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IL-18 associated with lung lymphoid aggregates drives IFN? production in severe COPD
Authors: E Briend, GJ Ferguson, M Mori, G Damera, K Stephenson, NA Karp, S Sethi, CK Ward, MA Sleeman, JS Erjefält, DK Finch
Respir. Res., 2017-08-22;18(1):159.
Species: Human
Sample Types: Cell Culture Supernates
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Production and regulation of interleukin-1 family cytokines at the materno-fetal interface
Authors: LM Scott, AH Bryant, A Rees, B Down, RH Jones, CA Thornton
Cytokine, 2017-07-13;0(0):.
Species: Human
Sample Types: Tissue Culture Supernates
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TB lymphadenitis is associated with enhanced baseline and antigen - specific induction of Type 1 and Type 17 cytokines and reduced IL-1? and IL-18 at the site of infection
Authors: GR Kathamuthu, K Moideen, D Baskaran, VV Banurekha, D Nair, G Sekar, R Sridhar, B Vidyajayan, G Gajendrara, DK Parandhama, A Srinivasan, S Babu
Clin. Vaccine Immunol, 2017-05-05;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Bortezomib treatment sensitizes oncolytic HSV-1 treated tumors to NK cell immunotherapy
Clin Cancer Res, 2016-07-07;0(0):.
Species: Mouse
Sample Types: Serum
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The Inflammasome and the Epidermal Growth Factor Receptor (EGFR) Are Involved in the Staphylococcus aureus-Mediated Induction of IL-1alpha and IL-1beta in Human Keratinocytes.
Authors: Simanski M, Rademacher F, Schroder L, Glaser R, Harder J
PLoS ONE, 2016-01-25;11(1):e0147118.
Species: Human
Sample Types: Cell Culture Supernates
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Angiopoietin-1 upregulates de novo expression of IL-1beta and Il1-Ra, and the exclusive release of Il1-Ra from human neutrophils.
Authors: Haddad L, Sirois M
PLoS ONE, 2014-02-12;9(2):e88980.
Species: Human
Sample Types: Cell Lysates
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Differential type 1 interferon-regulated gene expression in the brain during AIDS: interactions with viral diversity and neurovirulence.
Authors: Polyak M, Vivithanaporn P, Maingat F, Walsh J, Branton W, Cohen E, Meeker R, Power C
FASEB J, 2013-04-22;27(7):2829-44.
Species: Human
Sample Types: Cell Culture Supernates
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Interleukin-1alpha controls allergic sensitization to inhaled house dust mite via the epithelial release of GM-CSF and IL-33.
J. Exp. Med., 2012-07-16;209(8):1505-17.
Species: Human
Sample Types: Cell Culture Supernates
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Human epithelial cells establish direct antifungal defense through TLR4-mediated signaling.
Authors: Weindl G, Naglik JR, Kaesler S, Biedermann T, Hube B, Korting HC, Schaller M
J. Clin. Invest., 2007-12-01;117(12):3664-3672.
Species: Human
Sample Types: Cell Culture Supernates
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Pre-interleukin-1alpha expression reduces cell growth and increases interleukin-6 production in SaOS-2 osteosarcoma cells: Differential inhibitory effect of interleukin-1 receptor antagonist (icIL-1Ra1).
Authors: Palmer G, Trolliet S, Talabot-Ayer D, Mezin F, Magne D, Gabay C
Cytokine, 2005-07-21;31(2):153-60.
Species: Human
Sample Types: Cell Culture Supernates
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Fas and Fas-associated death domain protein regulate monocyte chemoattractant protein-1 expression by human smooth muscle cells through caspase- and calpain-dependent release of interleukin-1alpha.
Authors: Schaub FJ, Liles WC, Ferri N, Sayson K, Seifert RA, Bowen-Pope DF
Circ. Res., 2003-08-28;93(6):515-22.
Species: Human
Sample Types: Nuclear Extract
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Membrane-bound Interleukin-1 alpha mediates leukocyte adhesion during atherogenesis
Species: Human
Sample Types: Serum
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Very good kit, used on Human LiHep Plasma, 384-well plate format, sample used at 1/10 dilution and 4PL fitting.
Excellent ELISA.
We used on human serum with no dilution.
Excellent