Mouse LIX 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 CXCL5/LIX. 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: LIX
LPS-induced CXC chemokine (LIX) is a member of the murine chemokine family. It is 132 amino acids (aa) in length with a predicted molecular weight of 14 kDa prior to cleavage of the N-terminal signal peptide (aa 1–40). LIX has no true human ortholog but is closely related to human CXCL5/ENA-78 and CXCL6/GCP-2. It is widely expressed, and is secreted by multiple cell types including fibroblasts, thymic epithelium, platelets, vascular endothelium, and hepatocytes. LIX acts as a proinflammatory chemokine and is chemotactic for neutrophils.
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 LIX 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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Cooperation of immune regulators Tollip and surfactant protein A inhibits influenza A virus infection in mice
Authors: Schaunaman, N;Cervantes, D;Nichols, T;Numata, M;Ledford, JG;Kraft, M;Chu, HW;
Respiratory research
Species: Mouse
Sample Types: BALF
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Regulatory T cell-derived IL-1Ra suppresses the innate response to respiratory viral infection
Authors: Griffith, JW;Faustino, LD;Cottrell, VI;Nepal, K;Hariri, LP;Chiu, RS;Jones, MC;Julé, A;Gabay, C;Luster, AD;
Nature immunology
Species: Mouse
Sample Types: BALF
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Parkin Promotes Airway Inflammatory Response to Interferon Gamma
Authors: Dimasuay, KG;Schaunaman, N;Berg, B;Nichols, T;Chu, HW;
Biomedicines
Species: Mouse
Sample Types: Cell Culture Supernates
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Parkin promotes airway inflammatory response to interferon gamma
Authors: Dimasuay, KG;Berg, B;Schaunaman, N;Nichols, T;Chu, HW;
Research square
Species: Mouse
Sample Types: Cell Culture Supernates
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Role of Myeloid Cell-Specific TLR9 in Mitochondrial DNA-Induced Lung Inflammation in Mice
Authors: KG Dimasuay, B Berg, N Schaunaman, HW Chu
International Journal of Molecular Sciences, 2023-01-04;24(2):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Airway epithelial immunoproteasome subunit LMP7 protects against rhinovirus infection
Authors: KG Dimasuay, N Schaunaman, B Berg, D Cervantes, E Kruger, FL Heppner, DA Ferrington, HW Chu
Scientific Reports, 2022-08-25;12(1):14507.
Species: Mouse
Sample Types: Cell Lysates
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Local nebulization of 1alpha,25(OH)2D3 attenuates LPS-induced acute lung inflammation
Authors: J Serré, C Mathyssen, TT Ajime, T Heigl, L Verlinden, K Maes, A Verstuyf, D Cataldo, J Vanoirbeek, B Vanaudenae, W Janssens, G Gayan-Rami
Respiratory Research, 2022-03-29;23(1):76.
Species: Mouse
Sample Types: Tissue Homogenates
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Therapeutic senescence via GPCR activation in synovial fibroblasts facilitates resolution of arthritis
Authors: T Montero-Me, A Nagano, C Chelala, A Filer, CD Buckley, M Perretti
Nat Commun, 2020-02-06;11(1):745.
Species: Mouse
Sample Types: Tissue Homogenates
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SIRT3 promotes antimycobacterial defenses by coordinating mitochondrial and autophagic functions
Authors: TS Kim, YB Jin, YS Kim, S Kim, JK Kim, HM Lee, HW Suh, JH Choe, YJ Kim, BS Koo, HN Kim, M Jung, SH Lee, DK Kim, C Chung, JW Son, JJ Min, JM Kim, CX Deng, HS Kim, SR Lee, EK Jo
Autophagy, 2019-03-04;0(0):20-Jan.
Species: Mouse
Sample Types: Cell Culture Supernates
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The circadian regulator Bmal1 in joint mesenchymal cells regulates both joint development and inflammatory arthritis
Authors: LE Hand, SH Dickson, AJ Freemont, DW Ray, JE Gibbs
Arthritis Res. Ther., 2019-01-06;21(1):5.
Species: Mouse
Sample Types: Cell Culture Supernates
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p53 in Bronchial Club Cells Facilitates Chronic Lung Inflammation by Promoting Senescence
Authors: A Sagiv, A Bar-Shai, N Levi, M Hatzav, L Zada, Y Ovadya, L Roitman, G Manella, O Regev, J Majewska, E Vadai, R Eilam, SW Feigelson, M Tsoory, M Tauc, R Alon, V Krizhanovs
Cell Rep, 2018-03-27;22(13):3468-3479.
Species: Mouse
Sample Types: BALF
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Nlrp12 Mediates Adverse Neutrophil Recruitment during Influenza Virus Infection
Authors: EE Hornick, B Banoth, AM Miller, ZR Zacharias, N Jain, ME Wilson, KN Gibson-Cor, KL Legge, GA Bishop, FS Sutterwala, SL Cassel
J. Immunol., 2017-12-27;0(0):.
Species: Mouse
Sample Types: Tissue Homogenates
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Demyelination induced by oxidative stress is regulated by sphingosine 1-phosphate receptors
Authors: SA O'Sullivan, M Velasco-Es, KK Dev
Glia, 2017-04-04;0(0):.
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The time-of-day of myocardial infarction onset affects healing through oscillations in cardiac neutrophil recruitment
EMBO Mol Med, 2016-08-01;0(0):.
Species: Mouse
Sample Types: Plasma
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Matrix Metalloproteinase 9 Exerts Antiviral Activity against Respiratory Syncytial Virus.
Authors: Dabo A, Cummins N, Eden E, Geraghty P
PLoS ONE, 2015-08-18;10(8):e0135970.
Species: Mouse
Sample Types: BALF
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CXCR2-mediated tumor-associated neutrophil recruitment is regulated by IFN-beta.
Authors: Jablonska J, Wu C, Andzinski L, Leschner S, Weiss S
Int J Cancer, 2013-10-31;134(6):1346-58.
Species: Mouse
Sample Types: Tissue Homogenates
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Nucleosomes contribute to increase mesangial cell chemokine expression during the development of lupus nephritis.
Authors: Kanapathippillai P, Hedberg A, Fenton C, Fenton K
Cytokine, 2013-04-04;62(2):244-52.
Species: Mouse
Sample Types: Cell Culture Supernates
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Leukocyte ADAM17 Regulates Acute Pulmonary Inflammation.
Authors: Arndt PG, Strahan B, Wang Y, Long C, Horiuchi K, Walcheck B
PLoS ONE, 2011-05-16;6(5):e19938.
Species: Mouse
Sample Types: BALF
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IL-17A and TNF-alpha Exert Synergistic Effects on Expression of CXCL5 by Alveolar Type II Cells In Vivo and In Vitro.
Authors: Liu Y, Mei J, Gonzales L, Yang G, Dai N, Wang P, Zhang P, Favara M, Malcolm KC, Guttentag S, Worthen GS
J. Immunol., 2011-01-31;186(5):3197-205.
Species: Mouse
Sample Types: BALF
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Recombinant human tissue factor pathway inhibitor exerts anticoagulant, anti-inflammatory and antimicrobial effects in murine pneumococcal pneumonia.
Authors: Van Den Boogaard FE, Brands X, Schultz MJ, Levi M, Roelofs JJ, Van 't Veer C, van der Poll T
J. Thromb. Haemost., 2011-01-01;9(1):122-32.
Species: Mouse
Sample Types: BALF
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Absence or inhibition of matrix metalloproteinase-8 decreases ventilator-induced lung injury.
Authors: Albaiceta GM, Gutierrez-Fernandez A, Garcia-Prieto E, Puente XS, Parra D, Astudillo A, Campestre C, Cabrera S, Gonzalez-Lopez A, Fueyo A, Taboada F, Lopez-Otin C
Am. J. Respir. Cell Mol. Biol., 2009-12-07;43(5):555-63.
Species: Mouse
Sample Types: Tissue Homogenates
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Spatiotemporal expression of chemokines and chemokine receptors in experimental anti-myeloperoxidase antibody-mediated glomerulonephritis.
Authors: van der Veen BS, Petersen AH, Belperio JA, Satchell SC, Mathieson PW, Molema G, Heeringa P
Clin. Exp. Immunol., 2009-10-01;158(1):143-53.
Species: Mouse
Sample Types: Tissue Homogenates
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Allergic sensitization through the airway primes Th17-dependent neutrophilia and airway hyperresponsiveness.
Authors: Wilson RH, Whitehead GS, Nakano H, Free ME, Kolls JK, Cook DN
Am. J. Respir. Crit. Care Med., 2009-08-06;180(8):720-30.
Species: Mouse
Sample Types: BALF
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Matrix metalloproteinase-8 facilitates neutrophil migration through the corneal stromal matrix by collagen degradation and production of the chemotactic peptide Pro-Gly-Pro.
Authors: Lin M, Jackson P, Tester AM, Diaconu E, Overall CM, Blalock JE, Pearlman E
Am. J. Pathol., 2008-06-13;173(1):144-53.
Species: Mouse
Sample Types: Tissue Homogenates
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An essential role for IL-17 in preventing pathogen-initiated bone destruction: recruitment of neutrophils to inflamed bone requires IL-17 receptor-dependent signals.
Authors: Yu JJ, Ruddy MJ, Wong GC, Sfintescu C, Baker PJ, Smith JB, Evans RT, Gaffen SL
Blood, 2007-01-03;109(9):3794-802.
Species: Mouse
Sample Types: Serum
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Inhibition of angiogenesis by interleukin-4 gene therapy in rat adjuvant-induced arthritis.
Authors: Haas CS, Amin MA, Allen BB, Ruth JH, Haines GK, Woods JM, Koch AE
Arthritis Rheum., 2006-08-01;54(8):2402-14.
Species: Rat
Sample Types: Tissue Homogenates
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Matrix metalloproteinase-8 deficiency promotes granulocytic allergen-induced airway inflammation.
Authors: Gueders MM, Balbin M, Rocks N, Foidart JM, Gosset P, Louis R, Shapiro S, Lopez-Otin C, Noel A, Cataldo DD
J. Immunol., 2005-08-15;175(4):2589-97.
Species: Mouse
Sample Types: BALF
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Tissue inhibitor of metalloproteinase-1 deficiency amplifies acute lung injury in bleomycin-exposed mice.
Authors: Kim KH, Burkhart K, Chen P, Frevert CW, Randolph-Habecker J, Hackman RC, Soloway PD, Madtes DK
Am. J. Respir. Cell Mol. Biol., 2005-06-09;33(3):271-9.
Species: Mouse
Sample Types: BALF
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