Mouse 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 mouse 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 Mouse 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.
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Citations: Showing 1 - 10
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The IRAK1/IRF5 axis initiates IL-12 response by dendritic cells and control of Toxoplasma gondii infection
Authors: Pereira, M;Ramalho, T;Andrade, WA;Durso, DF;Souza, MC;Fitzgerald, KA;Golenbock, DT;Silverman, N;Gazzinelli, RT;
Cell reports
Species: Mouse
Sample Types: Serum, Cell Culture Supernates, Peritoneal Lavage Fluid
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Pexidartinib synergize PD-1 antibody through inhibiting treg infiltration by reducing TAM-derived CCL22 in lung adenocarcinoma
Authors: W Zhang, X Jiang, Y Zou, L Yuan, X Wang
Frontiers in Pharmacology, 2023-03-08;14(0):1092767.
Species: Mouse
Sample Types: Serum
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Combinatorial microRNA Loading into Extracellular Vesicles for Increased Anti-Inflammatory Efficacy
Authors: AE Pottash, D Levy, A Jeyaram, L Kuo, SM Kronstadt, W Chao, SM Jay
Non-coding RNA, 2022-10-21;8(5):.
Species: Mouse
Sample Types: Plasma
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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: Mouse
Sample Types: Cell Culture Supernates
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S100B Up-Regulates Macrophage Production of IL1beta and CCL22 and Influences Severity of Retinal Inflammation.
Authors: Niven J, Hoare J, McGowan D, Devarajan G, Itohara S, Gannage M, Teismann P, Crane I
PLoS ONE, 2015-07-23;10(7):e0132688.
Species: Mouse
Sample Types: Cell Culture Supernates
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Dipeptidylpeptidase 4 inhibition enhances lymphocyte trafficking, improving both naturally occurring tumor immunity and immunotherapy.
Authors: Barreira da Silva R, Laird M, Yatim N, Fiette L, Ingersoll M, Albert M
Nat Immunol, 2015-06-15;16(8):850-8.
Species: Mouse
Sample Types: Tissue Homogenates
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Network integration of parallel metabolic and transcriptional data reveals metabolic modules that regulate macrophage polarization.
Authors: Jha A, Huang S, Sergushichev A, Lampropoulou V, Ivanova Y, Loginicheva E, Chmielewski K, Stewart K, Ashall J, Everts B, Pearce E, Driggers E, Artyomov M
Immunity, 2015-03-17;42(3):419-30.
Species: Mouse
Sample Types: Cell Culture Supernates
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Influence of hypoxia-inducible factor 1-alpha on dendritic cell differentiation and migration.
Authors: Kohler T, Reizis B, Johnson RS, Weighardt H, Forster I
Eur. J. Immunol., 2012-05-01;42(5):1226-36.
Species: Mouse
Sample Types: Cell Culture Supernates
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Inflammatory chemokine release of astrocytes in vitro is reduced by all-trans retinoic acid.
Authors: van Neerven S, Regen T, Wolf D, Nemes A, Johann S, Beyer C, Hanisch UK, Mey J
J. Neurochem., 2010-06-16;114(5):1511-26.
Species: Mouse
Sample Types: Cell Culture Supernates
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Mycobacterium tuberculosis antigens specifically modulate CCR2 and MCP-1/CCL2 on lymphoid cells from human pulmonary hilar lymph nodes.
Authors: Arias MA, Jaramillo G, Lopez YP, Mejia N, Mejia C, Pantoja AE, Shattock RJ, Garcia LF, Griffin GE
J. Immunol., 2007-12-15;179(12):8381-91.
Species: Human
Sample Types: Cell Culture Supernates
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Immunosuppressive effects of CCL17 on pulmonary antifungal responses during pulmonary invasive aspergillosis.
Authors: Carpenter KJ, Hogaboam CM
Infect. Immun., 2005-11-01;73(11):7198-207.
Species: Mouse
Sample Types: Tissue Homogenates
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