Human CCL18/PARC 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 CCL18/PARC. 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: CCL18/PARC
Pulmonary and activation-regulated chemokine (PARC), also known as alternative macrophage activation-associated CC chemokine (AMAC)-1, macrophage inflammatory protein (MIP)-4, and dendritic cell chemokine (DC-CK1), is a novel CC chemokine that is highly homologous to MIP-1 alpha (61% amino acid sequence identity). In vitro, PARC mRNA expression is induced in alternatively activated macrophages by Th2 cytokines such as IL-4, IL-10 and IL-13, and inhibited by IFN-gamma.
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 CCL18/PARC 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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The IL-4/13-induced production of M2 chemokines by human lung macrophages is enhanced by adenosine and PGE2
Authors: Brollo, M;Salvator, H;Grassin-Delyle, S;Glorion, M;Descamps, D;Buenestado, A;Naline, E;Tenor, H;Tiotiu, A;Devillier, P;
International immunopharmacology
Species: Human
Sample Types: Cell Culture Supernates
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Prevention of M2 polarization and temporal limitation of differentiation in monocytes by extracellular ATP
Authors: Scherr, BF;Reiner, MF;Baumann, F;Höhne, K;Müller, T;Ayata, K;Müller-Quernheim, J;Idzko, M;Zissel, G;
BMC immunology
Species: Human
Sample Types: Cell Culture Supernates
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Onchocerca volvulus-specific antibody and cellular responses in onchocerciasis patients treated annually with ivermectin for 30 years and exposed to parasite transmission in central Togo
Authors: SI Johanns, RG Gantin, B Wangala, K Komlan, WA Halatoko, M Banla, P Karabou, AJ Luty, H Schulz-Key, C Köhler, PT Soboslay
PloS Neglected Tropical Diseases, 2022-05-03;16(5):e0010340.
Species: Human
Sample Types: Cell Culture Supernates
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Echinococcus multilocularis specific antibody, systemic cytokine, and chemokine levels, as well as antigen-specific cellular responses in patients with progressive, stable, and cured alveolar echinococcosis: A 10-year follow-up
Authors: B Grüner, L Peters, A Hillenbran, P Vo beta berg, J Schweiker, EG Rollmann, LH Rodriguez, J Blumhardt, S Burkert, P Kern, C Köhler, PT Soboslay
PloS Neglected Tropical Diseases, 2022-02-02;16(2):e0010099.
Species: Human
Sample Types: Cell Culture Supernates
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Tumor-Derived Extracellular Vesicles Induce CCL18 Production by Mast Cells: A Possible Link to Angiogenesis
Authors: I Shefler, P Salamon, T Zitman-Gal, YA Mekori
Cells, 2022-01-21;11(3):.
Species: Human
Sample Types: Cell Culture Supernates
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Insights into immunometabolism: A dataset correlating the 18FDG PET/CT maximum standard uptake value of the primary tumor with the CCL18 serum level in non-small cell lung cancer
Authors: T Dönmez, K Höhne, G Zissel, K Herrmann, H Hautzel, C Aigner, B Hegedüs, T Ploenes
Data in Brief, 2021-02-18;35(0):106859.
Species: Human
Sample Types: Serum
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Reprogramming of profibrotic macrophages for treatment of bleomycin-induced pulmonary fibrosis
Authors: F Zhang, EA Ayaub, B Wang, E Puchulu-Ca, YH Li, SU Hettiarach, SD Lindeman, Q Luo, S Rout, M Srinivasar, A Cox, K Tsoyi, C Nickerson-, IO Rosas, PS Low
EMBO Mol Med, 2020-06-29;0(0):e12034.
Species: Human
Sample Types: Cell Culture Supernates
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Genetic Variation in CCL18 Gene Influences CCL18 Expression and Correlates with Survival in Idiopathic Pulmonary Fibrosis-Part B
Authors: C Caliskan, B Seeliger, B Jäger, J Fuge, T Welte, O Terwolbeck, J Freise, CHM van Moorse, Y Zhang, A Prasse
J Clin Med, 2020-06-25;9(6):.
Species: Human
Sample Types: Serum
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Human interleukin-4-treated regulatory macrophages promote epithelial wound healing and reduce colitis in a mouse model
Authors: TS Jayme, G Leung, A Wang, ML Workentine, S Rajeev, A Shute, BE Callejas, N Mancini, PL Beck, R Panaccione, DM McKay
Sci Adv, 2020-06-05;6(23):eaba4376.
Species: Human
Sample Types: Cell Culture Supernates
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Pancreatic cancer-derived small extracellular vesical Ezrin regulates macrophage polarization and promotes metastasis
Authors: YT Chang, HY Peng, CM Hu, SC Huang, SC Tien, YM Jeng
Am J Cancer Res, 2020-01-01;10(1):12-37.
Species: Human
Sample Types: Cell Culture Supernates
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Intrapulmonary Autoantibodies to HSP72 Are Associated with Improved Outcomes in IPF
Authors: R Mills, A Mathur, LM Nicol, JJ Walker, AA Przybylski, AC Mackinnon, SEM Howie, WAH Wallace, I Dransfield, N Hirani
J Immunol Res, 2019-04-11;2019(0):1845128.
Species: Human
Sample Types: Cell Culture Supernates
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Prominent Levels of the Profibrotic Chemokine CCL18 during Peritonitis: In Vitro Downregulation by Vitamin D Receptor Agonists
Authors: M Ossorio, V Martínez, MA Bajo, G Del Peso, MJ Castro, S Romero, R Selgas, T Bellón
Biomed Res Int, 2018-04-04;2018(0):6415892.
Species: Human
Sample Types: Cell Culture Supernates
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Distinct Properties of Human M-CSF and GM-CSF Monocyte-Derived Macrophages to Simulate Pathological Lung Conditions In Vitro: Application to Systemic and Inflammatory Disorders with Pulmonary Involvement
Authors: A Lescoat, A Ballerie, Y Augagneur, C Morzadec, L Vernhet, O Fardel, P Jégo, S Jouneau, V Lecureur
Int J Mol Sci, 2018-03-17;19(3):.
Species: Human
Sample Types: Cell Culture Supernates
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Is serum level of CC chemokine ligand 18 a biomarker for the prediction of radiation induced lung toxicity (RILT)?
Authors: E Gkika, W Vach, S Adebahr, T Schimeck-J, A Brenner, TB Brunner, K Kaier, A Prasse, J Müller-Que, AL Grosu, G Zissel, U Nestle
PLoS ONE, 2017-09-28;12(9):e0185350.
Species: Human
Sample Types: Serum
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Image based Machine Learning for identification of macrophage subsets
Authors: HM Rostam, PM Reynolds, MR Alexander, N Gadegaard, AM Ghaemmagha
Sci Rep, 2017-06-14;7(1):3521.
Species: Human
Sample Types: Cell Culture Supernates
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Sustained low peritoneal effluent CCL18 levels are associated with preservation of peritoneal membrane function in peritoneal dialysis
Authors: M Ossorio, MA Bajo, G Del Peso, V Martínez, M Fernández, MJ Castro, A Rodríguez-, R Madero, T Bellón, R Selgas
PLoS ONE, 2017-04-17;12(4):e0175835.
Species: Human
Sample Types: Serum
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Validating glycoprotein non-metastatic melanoma B (gpNMB, osteoactivin), a new biomarker of Gaucher disease
Authors: Vagishwari Murugesan
Blood Cells Mol. Dis, 2016-12-13;0(0):.
Species: Human
Sample Types: Serum
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Exposure to diesel exhaust particle extracts (DEPe) impairs some polarization markers and functions of human macrophages through activation of AhR and Nrf2.
Authors: Jaguin M, Fardel O, Lecureur V
PLoS ONE, 2015-02-24;10(2):e0116560.
Species: Human
Sample Types: Cell Culture Supernates
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Macrophage activation in acute exacerbation of idiopathic pulmonary fibrosis.
Authors: Schupp J, Binder H, Jager B, Cillis G, Zissel G, Muller-Quernheim J, Prasse A
PLoS ONE, 2015-01-15;10(1):e0116775.
Species: Human
Sample Types: BALF
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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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Proteome-wide analysis and CXCL4 as a biomarker in systemic sclerosis
Authors: L van Bon, AJ Affandi, J Broen, RB Christmann, RJ Marijnisse, L Stawski, GA Farina, G Stifano, AL Mathes, M Cossu, M York, C Collins, M Wenink, R Huijbens, R Hesselstra, T Saxne, M DiMarzio, D Wuttge, SK Agarwal, JD Reveille, S Assassi, M Mayes, Y Deng, JP Drenth, J de Graaf, M den Heijer, CG Kallenberg, M Bijl, A Loof, WB van den Be, LA Joosten, V Smith, F de Keyser, R Scorza, C Lunardi, PL van Riel, M Vonk, W van Heerde, S Meller, B Homey, L Beretta, M Roest, M Trojanowsk, R Lafyatis, TR Radstake
N. Engl. J. Med, 2013-12-18;370(5):433-43.
Species: Human
Sample Types: Plasma
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Lung collagens perpetuate pulmonary fibrosis via CD204 and M2 macrophage activation.
Authors: Stahl M, Schupp J, Jager B, Schmid M, Zissel G, Muller-Quernheim J, Prasse A
PLoS ONE, 2013-11-20;8(11):e81382.
Species: Human
Sample Types: Cell Culture Supernates
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Serum level of CC-chemokine ligand 18 is increased in patients with non-small-cell lung cancer and correlates with survival time in adenocarcinomas.
Authors: Plones T, Krohn A, Burger M, Veelken H, Passlick B, Muller-Quernheim J, Zissel G
PLoS ONE, 2012-07-25;7(7):e41746.
Species: Human
Sample Types: Serum
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Potential biomarkers of osteonecrosis in Gaucher disease.
Authors: Pavlova EV, Deegan PB, Tindall J, McFarlane I, Mehta A, Hughes D, Wraith JE, Cox TM
Blood Cells Mol. Dis., 2010-11-13;46(1):27-33.
Species: Human
Sample Types: Serum
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Interferon-gamma reverses the immunosuppressive and protumoral properties and prevents the generation of human tumor-associated macrophages.
Authors: Duluc D, Corvaisier M, Blanchard S, Catala L, Descamps P, Gamelin E, Ponsoda S, Delneste Y, Hebbar M, Jeannin P
Int. J. Cancer, 2009-07-15;125(2):367-73.
Species: Human
Sample Types: Cell Culture Supernates
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Mature dendritic cells express functional thrombin receptors triggering chemotaxis and CCL18/pulmonary and activation-regulated chemokine induction.
Authors: Li X, Syrovets T, Paskas S, Laumonnier Y, Simmet T
J. Immunol., 2008-07-15;181(2):1215-23.
Species: Human
Sample Types: Cell Culture Supernates
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Inorganic arsenic activates reduced NADPH oxidase in human primary macrophages through a Rho kinase/p38 kinase pathway.
Authors: Lemarie A, Bourdonnay E, Morzadec C, Fardel O, Vernhet L
J. Immunol., 2008-05-01;180(9):6010-7.
Species: Human
Sample Types: Cell Culture Supernates
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Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells.
Authors: Duluc D, Delneste Y, Tan F, Moles MP, Grimaud L, Lenoir J, Preisser L, Anegon I, Catala L, Ifrah N, Descamps P, Gamelin E, Gascan H, Hebbar M, Jeannin P
Blood, 2007-09-11;110(13):4319-30.
Species: Human
Sample Types: Cell Culture Supernates
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CCL18 as an indicator of pulmonary fibrotic activity in idiopathic interstitial pneumonias and systemic sclerosis.
Authors: Prasse A, Pechkovsky DV, Toews GB, Schafer M, Eggeling S, Ludwig C, Germann M, Kollert F, Zissel G, Muller-Quernheim J
Arthritis Rheum., 2007-05-01;56(5):1685-93.
Species: Human
Sample Types: Serum
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High CCL18/PARC expression in articular cartilage and synovial tissue of patients with rheumatoid arthritis.
Authors: Momohara S, Okamoto H, Iwamoto T, Mizumura T, Ikari K, Kawaguchi Y, Takeuchi M, Kamatani N, Tomatsu T
J. Rheumatol., 2007-02-01;34(2):266-71.
Species: Human
Sample Types: Synovial Fluid
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Induction of prolonged infiltration of T lymphocytes and transient T lymphocyte-dependent collagen deposition in mouse lungs following adenoviral gene transfer of CCL18.
Authors: Luzina IG, Papadimitriou JC, Anderson R, Pochetuhen K, Atamas SP
Arthritis Rheum., 2006-08-01;54(8):2643-55.
Species: Human
Sample Types: BALF
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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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CCL18 is expressed in atopic dermatitis and mediates skin homing of human memory T cells.
Authors: Gunther C, Bello-Fernandez C, Kopp T, Kund J, Carballido-Perrig N, Hinteregger S, Fassl S, Schwarzler C, Lametschwandtner G, Stingl G, Biedermann T, Carballido JM
J. Immunol., 2005-02-01;174(3):1723-8.
Species: Human
Sample Types: Serum
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CC chemokine ligand 18, an atopic dermatitis-associated and dendritic cell-derived chemokine, is regulated by staphylococcal products and allergen exposure.
Authors: Pivarcsi A, Gombert M, Dieu-Nosjean MC, Kubitza R, Meller S, Rieker J, Muller A, Da Cunha L, Haahtela A, Sonkoly E, Fridman WH, Alenius H, Kemeny L, Ruzicka T, Zlotnik A
J. Immunol., 2004-11-01;173(9):5810-7.
Species: Human
Sample Types: Cell Culture Supernates
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