Human CCL22/MDC 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 CCL22/MDC. 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: CCL22/MDC
Macrophage-derived chemokine (MDC), also named stimulated T cell chemotactic protein (STCP-1) and ABCD-1, and now designated as CCL22, is a CC chemokine initially isolated from clones of monocyte-derived macrophages. At the amino acid sequence level, MDC shows less than 35% identity to other CC chemokine family members. Human MDC is expressed in dendritic cells, macrophages and activated monocytes. In addition, MDC expression is detected in thymus, lymph node and appendix tissues. At the amino acid sequence level, mouse and human MDC share 64% identity and 83% similarity.
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 CCL22/MDC 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.
25
Citations: Showing 1 - 10
Filter your results:
Filter by:
-
Neutrophil extracellular traps promote immunopathogenesis of virus-induced COPD exacerbations
Authors: Katsoulis, O;Toussaint, M;Jackson, MM;Mallia, P;Footitt, J;Mincham, KT;Meyer, GFM;Kebadze, T;Gilmour, A;Long, M;Aswani, AD;Snelgrove, RJ;Johnston, SL;Chalmers, JD;Singanayagam, A;
Nature communications
Species: Human
Sample Types: Sputum
-
Effect of Isoscopoletin on Cytokine Expression in HaCaT Keratinocytes and RBL-2H3 Basophils: Preliminary Study
Authors: Seo, DY;Park, JW;Kim, SH;Oh, SR;Han, SB;Kwon, OK;Ahn, KS;
International journal of molecular sciences
Species: Human
Sample Types: Cell Culture Supernates
-
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
-
Repeated exposure of bronchial epithelial cells to particular matter increases allergen-induced cytokine release and permeability
Authors: H Janbazacya, J van Bergen, S Varasteh, J Garssen, G Folkerts, S Braber
Cytokine, 2022-04-08;154(0):155878.
Species: Human
Sample Types: Cell Culture Supernates
-
IL-25 Induced ROS-Mediated M2 Macrophage Polarization via AMPK-Associated Mitophagy
Authors: ML Tsai, YG Tsai, YC Lin, YL Hsu, YT Chen, MK Tsai, WT Liao, YC Lin, CH Hung
International Journal of Molecular Sciences, 2021-12-21;23(1):.
Species: Human
Sample Types: Cell Culture Supernates
-
Skullcapflavone II Suppresses TNF-alpha/IFN-gamma-Induced TARC, MDC, and CTSS Production in HaCaT Cells
Authors: H Lee, DH Lee, JH Oh, JH Chung
International Journal of Molecular Sciences, 2021-06-16;22(12):.
Species: Human
Sample Types: Cell Culture Supernates
-
Head-to-head comparisons of Toxoplasma gondii and its near relative Hammondia hammondi reveal dramatic differences in the host response and effectors with species-specific functions
Authors: ZS Wong, SL Sokol-Borr, P Olias, JP Dubey, JP Boyle
PLoS Pathog., 2020-06-23;16(6):e1008528.
Species: Human
Sample Types: Cell Culture Supernates
-
Biomarkers and immunoprofiles associated with fetal abnormalities of ZIKV-positive pregnancies
Authors: SS Foo, W Chen, Y Chan, WS Lee, SA Lee, G Cheng, K Nielsen-Sa, P Brasil, JU Jung
JCI Insight, 2018-11-02;3(21):.
Species: Human
Sample Types: Serum
-
Rhododendron album Blume extract inhibits TNF-?/IFN-?-induced chemokine production via blockade of NF-?B and JAK/STAT activation in human epidermal keratinocytes
Authors: JW Park, HS Lee, Y Lim, JH Paik, OK Kwon, JH Kim, I Paryanto, P Yunianto, S Choi, SR Oh, KS Ahn
Int. J. Mol. Med., 2018-03-09;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
-
Human Placental Syncytiotrophoblasts Restrict Toxoplasma gondii Attachment and Replication and Respond to Infection by Producing Immunomodulatory Chemokines
Authors: SE Ander, EN Rudzki, N Arora, Y Sadovsky, CB Coyne, JP Boyle
MBio, 2018-01-09;9(1):.
Species: Human
Sample Types: Cell Culture Supernates
-
Forsythia suspensa Suppresses House Dust Mite Extract-Induced Atopic Dermatitis in NC/Nga Mice
PLoS ONE, 2016-12-09;11(12):e0167687.
Species: Human
Sample Types: Cell Culture Supernates
-
CCL22-specific T Cells: Modulating the immunosuppressive tumor microenvironment
Authors: Evelina Martinenai
Oncoimmunology, 2016-09-30;5(11):e1238541.
Species: Human
Sample Types: Cell Culture Supernates
-
Amyloid precursor protein mediated changes in intestinal epithelial phenotype in vitro.
Authors: Puig K, Manocha G, Combs C
PLoS ONE, 2015-03-05;10(3):e0119534.
Species: Human
Sample Types: Cell Culture Supernates
-
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
-
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
-
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
-
Selective activation of human dendritic cells by OM-85 through a NF-kB and MAPK dependent pathway.
Authors: Parola C, Salogni L, Vaira X, Scutera S, Somma P, Salvi V, Musso T, Tabbia G, Bardessono M, Pasquali C, Mantovani A, Sozzani S, Bosisio D
PLoS ONE, 2013-12-30;8(12):e82867.
Species: Human
Sample Types: Cell Culture Supernates
-
Identification of a New Phenotype of Tolerogenic Human Dendritic Cells Induced by Fungal Proteases from Aspergillus oryzae.
Authors: Zimmer A, Luce S, Gaignier F, Nony E, Naveau M, Biola-Vidamment A, Pallardy M, Van Overtvelt L, Mascarell L, Moingeon P
J. Immunol., 2011-03-02;186(7):3966-76.
Species: Human
Sample Types: Cell Culture Supernates
-
IRF5 promotes inflammatory macrophage polarization and T(H)1-T(H)17 responses.
Authors: Krausgruber T, Blazek K, Smallie T
Nat. Immunol., 2011-01-16;12(3):231-8.
Species: Human
Sample Types: Cell Culture Supernates
-
Divergent effects of hypoxia on dendritic cell functions.
Authors: Mancino A, Schioppa T, Larghi P, Pasqualini F, Nebuloni M, Chen IH, Sozzani S, Austyn JM, Mantovani A, Sica A
Blood, 2008-08-11;112(9):3723-34.
Species: Human
Sample Types: Cell Culture Supernates
-
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
-
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: Whole Cells
-
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
-
Ligation of the FcR gamma chain-associated human osteoclast-associated receptor enhances the proinflammatory responses of human monocytes and neutrophils.
Authors: Merck E, Gaillard C, Scuiller M, Scapini P, Cassatella MA, Trinchieri G, Bates EE
J. Immunol., 2006-03-01;176(5):3149-56.
Species: Human
Sample Types: Cell Culture Supernates
-
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
FAQs
No product specific FAQs exist for this product, however you may
View all ELISA FAQsReviews for Human CCL22/MDC DuoSet ELISA
Average Rating: 4 (Based on 1 Review)
Have you used Human CCL22/MDC DuoSet ELISA?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
Filter by: