Mouse CXCL9/MIG 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 CXCL9/MIG. 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: CXCL9/MIG
CXCL9, also known as MIG, is a chemokine that is induced in macrophages and in CNS glial cells in response to IFN-gamma. CXCL9 signals through CXCR3 to induce the chemoattraction of activated T cells and tumor-infiltrating lymphocytes.
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 CXCL9/MIG 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.
22
Citations: Showing 1 - 10
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Poly I:C vaccination drives transient CXCL9 expression near B cell follicles in the lymph node through type-I and type-II interferon signaling
Authors: Ball, AG;Morgaenko, K;Anbaei, P;Ewald, SE;Pompano, RR;
Cytokine
Species: Mouse
Sample Types: Tissue Supernates
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TBK1 and IKK? protect target cells from IFN?-mediated T cell killing via an inflammatory apoptotic mechanism
Authors: Sun, ND;Carr, AR;Krogman, EN;Chawla, Y;Zhong, J;Guttormson, MC;Chan, M;Hsu, MA;Dong, H;Bogunovic, D;Pandey, A;Rogers, LM;Ting, AT;
bioRxiv : the preprint server for biology
Species: Mouse
Sample Types: Cell Culture Supernates
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Reduction of chemokine CXCL9 expression by omega-3 fatty acids via ADP-ribosylhydrolase ARH3 in MIN6 insulin-producing cells
Authors: You, Y;Sarkar, S;Deiter, C;Elliott, EC;Nicora, CD;Mirmira, RG;Sussel, L;Nakayasu, ES;
bioRxiv : the preprint server for biology
Species: Mouse
Sample Types: Cell Culture Supernates
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A novel aptamer-based small RNA delivery platform and its application to cancer therapy
Authors: Tanno, T;Zhang, P;Bailey, C;Wang, Y;Ittiprasert, W;Devenport, M;Zheng, P;Liu, Y;
Genes & diseases
Species: Mouse, Transgenic Mouse
Sample Types: Plasma
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Ionizing Radiation Selectively Increases CXC Ligand 10 Level via the DNA-Damage-Induced p38 MAPK-STAT1 Pathway in Murine J774A.1 Macrophages
Authors: YN Seo, JS Baik, SM Lee, JE Lee, HR Ahn, MS Lim, MT Park, SD Kim
Cells, 2023-03-25;12(7):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Type I IFN stimulates lymph node stromal cells from adult and old mice during a West Nile virus infection
Authors: AK Bennett, M Richner, MD Mun, JM Richner
bioRxiv : the preprint server for biology, 2023-01-05;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Bronchoalveolar lavage fluid reveals factors contributing to the efficacy of PD-1 blockade in lung cancer
Authors: K Masuhiro, M Tamiya, K Fujimoto, S Koyama, Y Naito, A Osa, T Hirai, H Suzuki, N Okamoto, T Shiroyama, K Nishino, Y Adachi, T Nii, Y Kinugasa-K, A Kajihara, T Morita, S Imoto, S Uematsu, T Irie, D Okuzaki, T Aoshi, Y Takeda, T Kumagai, T Hirashima, A Kumanogoh
JCI Insight, 2022-05-09;7(9):.
Species: Mouse
Sample Types: Tissue Homogenates
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CXCL9 inhibits tumour growth and drives anti-PD-L1 therapy in ovarian cancer
Authors: S Seitz, TF Dreyer, C Stange, K Steiger, R Bräuer, L Scheutz, G Multhoff, W Weichert, M Kiechle, V Magdolen, H Bronger
British Journal of Cancer, 2022-03-21;0(0):.
Species: Mouse
Sample Types: Cell culture supernate
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Oxidized carbon black nanoparticles induce endothelial damage through C-X-C chemokine receptor 3-mediated pathway
Authors: N Majumder, M Velayutham, D Bitounis, VK Kodali, MH Hasan Mazu, J Amedro, VV Khramtsov, A Erdely, T Nurkiewicz, P Demokritou, EE Kelley, S Hussain
Redox Biology, 2021-10-04;47(0):102161.
Species: Mouse
Sample Types: BALF
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NK cell recruitment limits tissue damage during an enteric helminth infection
Authors: ME Gentile, Y Li, A Robertson, K Shah, G Fontes, E Kaufmann, B Polese, N Khan, M Parisien, HM Munter, JN Mandl, L Diatchenko, M Divangahi, IL King
Mucosal Immunol, 2019-11-27;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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LIF regulates CXCL9 in tumor-associated macrophages and prevents CD8+ T cell tumor-infiltration impairing anti-PD1 therapy
Authors: M Pascual-Ga, E Bonfill-Te, E Planas-Rig, C Rubio-Pere, R Iurlaro, A Arias, I Cuartas, A Sala-Hojma, L Escudero, F Martínez-R, I Huber-Ruan, P Nuciforo, L Pedrosa, C Marques, I Braña, E Garralda, M Vieito, M Squatrito, E Pineda, F Graus, C Espejo, J Sahuquillo, J Tabernero, J Seoane
Nat Commun, 2019-06-11;10(1):2416.
Species: Mouse
Sample Types: Cell Culture Supernates
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In-depth characterization of congenital Zika syndrome in immunocompetent mice: Antibody-dependent enhancement and an antiviral peptide therapy
Authors: VN Camargos, G Foureaux, DC Medeiros, VT da Silveir, CM Queiroz-Ju, ALB Matosinhos, AFA Figueiredo, CDF Sousa, TP Moreira, VF Queiroz, ACF Dias, KTO Santana, I Passos, ALCV Real, LC Silva, FAG Mourão, NT Wnuk, MAP Oliveira, S Macari, T Silva, GP Garlet, JA Jackman, FM Soriani, MFD Moraes, EMAM Mendes, FM Ribeiro, GMJ Costa, AL Teixeira, NJ Cho, ACP Oliveira, MM Teixeira, VV Costa, DG Souza
EBioMedicine, 2019-05-23;0(0):.
Species: Mouse
Sample Types: Plasma
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The Shc1 adaptor simultaneously balances Stat1 and Stat3 activity to promote breast cancer immune suppression
Authors: R Ahn, V Sabourin, AM Bolt, S Hébert, S Totten, N De Jay, MC Festa, YK Young, YK Im, T Pawson, AE Koromilas, WJ Muller, KK Mann, CL Kleinman, J Ursini-Sie
Nat Commun, 2017-03-09;8(0):14638.
Species: Mouse
Sample Types: Cell Culture Supernates
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NOD2 regulates CXCR3-dependent CD8+ T cell accumulation in intestinal tissues with acute injury.
Authors: Wu X, Lahiri A, Haines G, Flavell R, Abraham C
J Immunol, 2014-03-03;192(7):3409-18.
Species: Mouse
Sample Types: Cell Culture Supernates
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CD3(-)NK1.1(+) cells aid in the early induction of a Th1 response to an attaching and effacing enteric pathogen.
Authors: Reid-Yu S, Small C, Coombes B
Eur J Immunol, 2013-07-12;43(10):2638-49.
Species: Mouse
Sample Types: Tissue Homogenates
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IL-10 mediated regulation of liver inflammation during acute murine cytomegalovirus infection.
Authors: Gaddi, Pamela J, Crane, Meredith, Kamanaka, Masahito, Flavell, Richard, Yap, George S, Salazar-Mather, Thais P
PLoS ONE, 2012-08-03;7(8):e42850.
Species: Mouse
Sample Types: Serum
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Phosphoinositide 3-kinase gamma plays a critical role in bleomycin-induced pulmonary inflammation and fibrosis in mice.
Authors: Russo RC, Garcia CC, Barcelos LS, Rachid MA, Guabiraba R, Roffe E, Souza AL, Sousa LP, Mirolo M, Doni A, Cassali GD, Pinho V, Locati M, Teixeira MM
J. Leukoc. Biol., 2010-11-02;89(2):269-82.
Species: Mouse
Sample Types: Tissue Homogenates
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Th17 cells are the dominant T cell subtype primed by Shigella flexneri mediating protective immunity.
Authors: Sellge G, Magalhaes JG, Konradt C, Fritz JH, Salgado-Pabon W, Eberl G, Bandeira A, Di Santo JP, Sansonetti PJ, Phalipon A
J. Immunol., 2010-01-20;184(4):2076-85.
Species: Mouse
Sample Types: Cell Culture Supernates
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Toll-like receptor (TLR)2 and TLR3 synergy and cross-inhibition in murine myeloid dendritic cells.
Authors: Vanhoutte F, Paget C, Breuilh L, Fontaine J, Vendeville C, Goriely S, Ryffel B, Faveeuw C, Trottein F
Immunol. Lett., 2007-12-17;116(1):86-94.
Species: Mouse
Sample Types: Cell Culture Supernates
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CXCR3-dependent recruitment of antigen-specific T lymphocytes to the liver during murine cytomegalovirus infection.
Authors: Hokeness KL, Deweerd ES, Munks MW, Lewis CA, Gladue RP, Salazar-Mather TP
J. Virol., 2006-11-15;81(3):1241-50.
Species: Mouse
Sample Types: Cell Culture Supernates
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CXCR3 and IFN protein-10 in Pneumocystis pneumonia.
Authors: McAllister F, Ruan S, Steele C, Zheng M, McKinley L, Ulrich L, Marrero L, Shellito JE, Kolls JK
J. Immunol., 2006-08-01;177(3):1846-54.
Species: Mouse
Sample Types: Tissue Homogenates
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Negative regulation of eosinophil recruitment to the lung by the chemokine monokine induced by IFN-gamma (Mig, CXCL9).
Authors: Fulkerson PC, Zimmermann N, Brandt EB, Muntel EE, Doepker MP, Kavanaugh JL, Mishra A, Witte DP, Zhang H, Farber JM, Yang M, Foster PS, Rothenberg ME
Proc. Natl. Acad. Sci. U.S.A., 2004-02-09;101(7):1987-92.
Species: Mouse
Sample Types: BALF
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