Human 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 human 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 Human 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.
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Citations: Showing 1 - 10
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Myocardial transcriptomic analysis of diabetic patients with aortic stenosis: key role for mitochondrial calcium signaling
Authors: Cherpaz, M;Meugnier, E;Seillier, G;Pozzi, M;Pierrard, R;Leboube, S;Farhat, F;Vola, M;Obadia, JF;Amaz, C;Chalabreysse, L;May, C;Chanon, S;Brun, C;Givre, L;Bidaux, G;Mewton, N;Derumeaux, G;Bergerot, C;Paillard, M;Thibault, H;
Cardiovascular diabetology
Species: Human
Sample Types: Serum
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Plasma markers in pulmonary hypertension subgroups correlate with patient survival
Authors: T Koudstaal, D van Uden, JAC van Hulst, P Heukels, IM Bergen, LW Geenen, VJM Baggen, AE van den Bo, LM van den To, PP Chandoesin, M Kool, E Boersma, RW Hendriks, KA Boomars
Respiratory Research, 2021-05-04;22(1):137.
Species: Human
Sample Types: Plasma
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Adipocytokines in Untreated Newly Diagnosed Rheumatoid Arthritis: Association with Circulating Chemokines and Markers of Inflammation
Authors: GK Vasileiadi, AC Lundell, Y Zhang, K Andersson, I Gjertsson, A Rudin, C Maglio
Biomolecules, 2021-02-21;11(2):.
Species: Human
Sample Types: Plasma
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Clinical Response to the CD95-Ligand Inhibitor Asunercept Is Defined by a Pro-Inflammatory Serum Cytokine Profile
Authors: A Radujkovic, T Boch, F Nolte, D Nowak, C Kunz, A Gieffers, C Müller-Tid, P Dreger, WK Hofmann, T Luft
Cancers, 2020-12-08;12(12):.
Species: Human
Sample Types: Serum
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Interleukin-18 and Hematopoietic Recovery after Allogeneic Stem Cell Transplantation
Authors: A Radujkovic, L Kordelas, R Bogdanov, C Müller-Tid, DW Beelen, P Dreger, T Luft
Cancers (Basel), 2020-09-28;12(10):.
Species: Human
Sample Types: Serum
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Human BCL-G regulates secretion of inflammatory chemokines but is dispensable for induction of apoptosis by IFN-gamma and TNF-alpha in intestinal epithelial cells
Authors: JA Woznicki, P Flood, M Bustamante, P Stamou, G Moloney, A Fanning, SA Zulquernai, J McCarthy, F Shanahan, S Melgar, K Nally
Cell Death Dis, 2020-01-27;11(1):68.
Species: Human
Sample Types: Cell Culture Supernates
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The interferon-gamma pathway is selectively up-regulated in the liver of patients with secondary hemophagocytic lymphohistiocytosis
Authors: G Prencipe, C Bracaglia, I Caiello, A Pascarella, P Francalanc, M Pardeo, A Meneghel, G Martini, MN Rossi, A Insalaco, G Marucci, V Nobili, M Spada, F Zulian, F De Benedet
PLoS ONE, 2019-12-17;14(12):e0226043.
Species: Human
Sample Types: Plasma
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IL-27 Modulates Chemokine Production in TNF-? -Stimulated Human Oral Epithelial Cells
Authors: Y Hosokawa, I Hosokawa, K Ozaki, T Matsuo
Cell. Physiol. Biochem., 2017-10-05;43(3):1198-1206.
Species: Human
Sample Types: Cell Culture Supernates
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CXCR3 mediates ascites-directed tumor cell migration and predicts poor outcome in ovarian cancer patients
Authors: C Windmüller, D Zech, S Avril, M Boxberg, T Dawidek, B Schmalfeld, M Schmitt, M Kiechle, H Bronger
Oncogenesis, 2017-05-15;6(5):e331.
Species: Human
Sample Types: Cell Lysates
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Identification of an immunogenic neo-epitope encoded by mouse sarcoma using CXCR3 ligand mRNAs as sensors
Authors: K Fujii, Y Miyahara, N Harada, D Muraoka, M Komura, R Yamaguchi, H Yagita, J Nakamura, S Sugino, S Okumura, S Imoto, S Miyano, H Shiku
Oncoimmunology, 2017-03-20;6(5):e1306617.
Species: Human
Sample Types: Cell Culture Supernates
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Suction blistering the lesional skin of vitiligo patients reveals useful biomarkers of disease activity
Authors: JP Strassner, M Rashighi, M Ahmed Refa, JM Richmond, JE Harris
J. Am. Acad. Dermatol, 2017-03-01;0(0):.
Species: Human
Sample Types: Blister Fluid
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CXCL9 and CXCL10 predict survival and are regulated by cyclooxygenase inhibition in advanced serous ovarian cancer
Br J Cancer, 2016-08-04;0(0):.
Species: Human
Sample Types: Tissue Homogenates
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PRC2 Epigenetically Silences Th1-Type Chemokines to Suppress Effector T-Cell Trafficking in Colon Cancer.
Authors: Nagarsheth N, Peng D, Kryczek I, Wu K, Li W, Zhao E, Zhao L, Wei S, Frankel T, Vatan L, Szeliga W, Dou Y, Owens S, Marquez V, Tao K, Huang E, Wang G, Zou W
Cancer Res, 2015-11-13;76(2):275-82.
Species: Human
Sample Types: Cell Culture Supernates
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Effect of JAK Inhibitors on Release of CXCL9, CXCL10 and CXCL11 from Human Airway Epithelial Cells.
Authors: Fenwick P, Macedo P, Kilty I, Barnes P, Donnelly L
PLoS ONE, 2015-06-19;10(6):e0128757.
Species: Human
Sample Types: Cell Culture Supernates
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Human B cells induce dendritic cell maturation and favour Th2 polarization by inducing OX-40 ligand.
Authors: Maddur M, Sharma M, Hegde P, Stephen-Victor E, Pulendran B, Kaveri S, Bayry J
Nat Commun, 2014-06-09;5(0):4092.
Species: Human
Sample Types: Cell Culture Supernates
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CXCR3 axis in patients with primary biliary cirrhosis: a possible novel mechanism of the effect of ursodeoxycholic acid.
Authors: Manousou P, Kolios G, Drygiannakis I, Koulentaki M, Pyrovolaki K, Voumvouraki A, Notas G, Bourikas L, Papadaki H, Kouroumalis E
Clin Exp Immunol, 2013-04-01;172(1):9-15.
Species: Human
Sample Types: Serum
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Abnormalities in serum chemokine levels in euthymic patients with bipolar disorder.
Authors: Brietzke E, Kauer-Sant'Anna M, Teixeira AL, Kapczinski F
Brain Behav. Immun., 2009-05-04;23(8):1079-82.
Species: Human
Sample Types: Serum
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Increased serum levels of CCL11/eotaxin in schizophrenia.
Authors: Teixeira AL, Reis HJ, Nicolato R, Brito-Melo G, Correa H, Teixeira MM, Romano-Silva MA
Prog. Neuropsychopharmacol. Biol. Psychiatry, 2007-11-23;32(3):710-4.
Species: Human
Sample Types: Serum
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The role of IkappaB kinase 2, but not activation of NF-kappaB, in the release of CXCR3 ligands from IFN-gamma-stimulated human bronchial epithelial cells.
Authors: Tudhope SJ, Catley MC, Fenwick PS, Russell RE, Rumsey WL, Newton R, Barnes PJ, Donnelly LE
J. Immunol., 2007-11-01;179(9):6237-45.
Species: Human
Sample Types: Cell Culture Supernates
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Modification of TLR-induced activation of human dendritic cells by type I IFN: synergistic interaction with TLR4 but not TLR3 agonists.
Authors: Walker J, Tough DF
Eur. J. Immunol., 2006-07-01;36(7):1827-36.
Species: Human
Sample Types: Cell Culture Supernates
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Diesel exhaust exposure favors TH2 cell recruitment in nonatopic subjects by differentially regulating chemokine production.
Authors: Chang Y, Senechal S, de Nadai P, Chenivesse C, Gilet J, Vorng H, Legendre B, Tonnel AB, Wallaert B, Lassalle P, Tsicopoulos A
J. Allergy Clin. Immunol., 2006-06-21;118(2):354-60.
Species: Human
Sample Types: Cell Culture Supernates
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CXCR 3 activation promotes lymphocyte transendothelial migration across human hepatic endothelium under fluid flow.
Authors: Curbishley SM, Eksteen B, Gladue RP, Lalor P, Adams DH
Am. J. Pathol., 2005-09-01;167(3):887-99.
Species: Human
Sample Types: Cell Culture Supernates
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Prediction of acute renal allograft rejection by urinary monokine induced by IFN-gamma (MIG).
Authors: Hauser IA, Spiegler S, Kiss E, Gauer S, Sichler O, Scheuermann EH, Ackermann H, Pfeilschifter JM, Geiger H, Grone HJ, Radeke HH
J. Am. Soc. Nephrol., 2005-04-27;16(6):1849-58.
Species: Human
Sample Types: Urine
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Chemokines and chemokine receptors in inflammatory demyelinating neuropathies: a central role for IP-10.
Authors: Kieseier BC, #N*´)2ZP, Tani M, y§/ÔcØÞ, Mahad D, Oka N, Ho T, Woodroofe N, Griffin JW, Toyka KV, Ransohoff RM, Hartung HP
2002-04-01;125(0):823-34.
Species: Human
Sample Types: CSF
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