Mouse IL-28A/B (IFN-lambda 2/3) 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-28A/B (IFN-lambda 2/3). 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-28A/B (IFN-lambda 2/3)
Human/mouse IL-28A (IFN-lambda 2), IL-28B (IFN-lambda 3), and human IL-29 (IFN-lambda 1) are class II cytokine receptor ligands that are distantly related to members of the IL-10 family (11 - 13% amino acid (aa) sequence identity) and type I IFN family (15 - 19% aa sequence identity). These cytokines exert bioactivities that overlap those of type I IFNs, including antiviral activity and up-regulation of MHC class I antigen expression. The proteins signal through the same heterodimeric receptor complex that is composed of the IL-10 receptor beta (IL-10 R beta) and a novel IL-28 receptor alpha (IL-28 R alpha, also known as IFN-lambda R1). Mouse IL-28 shares 61%, 62%, and 52% aa identity with human IL-28A, IL-28B, and IL-29, respectively.
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-28A/B (IFN-lambda 2/3) 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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IFN-?3 is induced by Leishmania donovani and can inhibit parasite growth in cell line models but not in the mouse model, while it shows a significant association with leishmaniasis in humans
Authors: De, M;Sukla, S;Bharatiya, S;Keshri, S;Roy, DG;Roy, S;Dutta, D;Saha, S;Ejazi, SA;Ravichandiran, V;Ali, N;Chatterjee, M;Chinnaswamy, S;
Infection and immunity
Species: Mouse
Sample Types: Cell Culture Supernates
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Bacterial-induced or passively administered interferon gamma conditions the lung for early control of SARS-CoV-2
Authors: Hilligan, KL;Namasivayam, S;Clancy, CS;Baker, PJ;Old, SI;Peluf, V;Amaral, EP;Oland, SD;O'Mard, D;Laux, J;Cohen, M;Garza, NL;Lafont, BAP;Johnson, RF;Feng, CG;Jankovic, D;Lamiable, O;Mayer-Barber, KD;Sher, A;
Nature communications
Species: Mouse
Sample Types: Tissue Homogenates
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Nei-like DNA glycosylase 2 (NEIL2) selectively antagonizes interferon-beta expression upon respiratory syncytial virus infection
Authors: Pan, L;Xue, Y;Wang, K;Zheng, X;Islam, A;Tapryal, N;Chakraborty, A;Bacsi, A;Ba, X;Hazra, TK;Boldogh, I;
The Journal of biological chemistry
Species: Mice
Sample Types: BALF
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IL-20 Cytokines Are Involved in Epithelial Lesions Associated with Virus-Induced COPD Exacerbation in Mice
Authors: M Le Roux, A Ollivier, G Kervoaze, T Beke, L Gillet, M Pichavant, P Gosset
Biomedicines, 2021-12-05;9(12):.
Species: Mouse
Sample Types: BALF
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Murine Type III interferons are functionally redundant and correlate with bacterial burden during influenza/bacterial super-infection
Authors: HE Rich, D Antos, CC McCourt, WQ Zheng, LJ Devito, KJ McHugh, R Gopal, J Wang, JF Alcorn
PLoS ONE, 2021-10-07;16(10):e0255309.
Species: Mouse
Sample Types: BALF
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Intestinal antiviral signaling is controlled by autophagy gene Epg5 independent of the microbiota
Authors: S Lee, G Kalugotla, H Ingle, R Rodgers, C Wu, Y Wang, Y Li, X Yang, J Zhang, NR Borella, H Deng, L Droit, R Hill, ST Peterson, C Desai, D Lawrence, Q Lu, MT Baldridge
Autophagy, 2021-09-14;0(0):1-16.
Species: Mouse
Sample Types: Serum
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A non-neutralizing antibody broadly protects against influenza virus infection by engaging effector cells
Authors: YA Ko, YH Yu, YF Wu, YC Tseng, CL Chen, KS Goh, HY Liao, TH Chen, TR Cheng, AS Yang, CH Wong, C Ma, KI Lin
PloS Pathogens, 2021-08-05;17(8):e1009724.
Species: Mouse
Sample Types: Tissue Homogenates
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DHX15 is required to control RNA virus-induced intestinal inflammation
Authors: J Xing, X Zhou, M Fang, E Zhang, LJ Minze, Z Zhang
Cell Reports, 2021-06-22;35(12):109205.
Species: Mouse
Sample Types: Cell Culture Supernates, Tissue Homogenates
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Murine astrovirus tropism for goblet cells and enterocytes facilitates an IFN-&lambda response in vivo and in enteroid cultures
Authors: H Ingle, E Hassan, J Gawron, B Mihi, Y Li, EA Kennedy, G Kalugotla, H Makimaa, S Lee, P Desai, KG McDonald, MS Diamond, RD Newberry, M Good, MT Baldridge
Mucosal Immunology, 2021-03-05;0(0):.
Species: Mouse
Sample Types: Serum
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Interferon lambda promotes immune dysregulation and tissue inflammation in TLR7-induced lupus
Authors: RR Goel, X Wang, LJ O'Neil, S Nakabo, K Hasneen, S Gupta, G Wigerblad, LP Blanco, JB Kopp, MI Morasso, SV Kotenko, ZX Yu, C Carmona-Ri, MJ Kaplan
Proc. Natl. Acad. Sci. U.S.A., 2020-02-24;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates, Serum
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Distinct roles for MDA5 and TLR3 in the acute response to inhaled double-stranded RNA
Authors: JM Veazey, TJ Chapman, TR Smyth, SE Hillman, SI Eliseeva, SN Georas
PLoS ONE, 2019-05-08;14(5):e0216056.
Species: Mouse
Sample Types: BALF fluid
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Corticosteroid suppression of antiviral immunity increases bacterial loads and mucus production in COPD exacerbations
Authors: A Singanayag, N Glanville, JL Girkin, YM Ching, A Marcellini, JD Porter, M Toussaint, RP Walton, LJ Finney, J Aniscenko, J Zhu, MB Trujillo-T, MA Calderazzo, C Grainge, SL Loo, PC Veerati, PS Pathinayak, KS Nichol, AT Reid, PL James, R Solari, PAB Wark, DA Knight, MF Moffatt, WO Cookson, MR Edwards, P Mallia, NW Bartlett, SL Johnston
Nat Commun, 2018-06-08;9(1):2229.
Species: Mouse
Sample Types: BALF
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Interferon-? Mediates Non-redundant Front-Line Antiviral Protection against Influenza Virus Infection without Compromising Host Fitness
Authors: IE Galani, V Triantafyl, EE Eleminiado, O Koltsida, A Stavropoul, M Manioudaki, D Thanos, SE Doyle, SV Kotenko, K Thanopoulo, E Andreakos
Immunity, 2017-05-16;46(5):875-890.e6.
Species: Mouse
Sample Types: Cell Culture Supernates
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TRAIL signaling is pro-inflammatory and pro-viral in a murine model of rhinovirus 1B infection
Authors: Joerg Mattes
Am. J. Physiol. Lung Cell Mol. Physiol., 2016-11-11;0(0):ajplung.00200.
Species: Mouse
Sample Types: Tissue Homogenates
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Identification of Toll-like receptor 9 as parapoxvirus ovis-sensing receptor in plasmacytoid dendritic cells.
Authors: von Buttlar H, Siegemund S, Buttner M, Alber G
PLoS ONE, 2014-08-29;9(8):e106188.
Species: Mouse
Sample Types: Cell Culture Supernates
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IRF-3, IRF-7, and IPS-1 promote host defense against acute human metapneumovirus infection in neonatal mice.
Authors: Spann K, Loh Z, Lynch J, Ullah A, Zhang V, Baturcam E, Werder R, Khajornjiraphan N, Rudd P, Loo Y, Suhrbier A, Gale M, Upham J, Phipps S
Am J Pathol, 2014-04-13;184(6):1795-806.
Species: Mouse
Sample Types: Cell Lysates
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IPS-1 is essential for type III IFN production by hepatocytes and dendritic cells in response to hepatitis C virus infection.
Authors: Okamoto, Masaaki, Oshiumi, Hiroyuki, Azuma, Masahiro, Kato, Nobuyuki, Matsumoto, Misako, Seya, Tsukasa
J Immunol, 2014-02-14;192(6):2770-7.
Species: Mouse
Sample Types:
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IL-15 complexes induce NK- and T-cell responses independent of type I IFN signaling during rhinovirus infection.
Authors: Jayaraman A, Jackson D, Message S, Pearson R, Aniscenko J, Caramori G, Mallia P, Papi A, Shamji B, Edwards M, Westwick J, Hansel T, Stanciu L, Johnston S, Bartlett N
Mucosal Immunol, 2014-01-29;7(5):1151-64.
Species: Mouse
Sample Types: BALF
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Type I and type III interferons drive redundant amplification loops to induce a transcriptional signature in influenza-infected airway epithelia.
Authors: Crotta S, Davidson S, Mahlakoiv T, Desmet C, Buckwalter M, Albert M, Staeheli P, Wack A
PLoS Pathog, 2013-11-21;9(11):e1003773.
Species: Mouse
Sample Types: Cell Culture Supernates
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The IFN regulatory factor 7-dependent type I IFN response is not essential for early resistance against murine cytomegalovirus infection.
Authors: Steinberg C, Eisenacher K, Gross O, Reindl W, Schmitz F, Ruland J, Krug A
Eur. J. Immunol., 2009-04-01;39(4):1007-18.
Species: Mouse
Sample Types: Serum
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