Human CCL17/TARC DuoSet ELISA DY364: R&D Systems

Human CCL17/TARC DuoSet ELISA

(10 citations)   
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Sample Type & Volume Required
    100 µL
  • Range
    7.81 - 500 pg/mL
  • Sufficient Materials
    For fifteen 96-well plates*
  • Specificity
    Please see the product datasheet
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
Ancillary Reagent Kit Available

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CCL17/TARC. 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)

Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CCL17/TARC
CCL17/TARC is a chemokine that interacts with CCR4 and CCR8 to induce the chemoattraction of activated Th2 cells, basophils, and NK cells. It is produced by thymic dendritic cells, lymph node dendritic cells, monocytes, CD4+ T cells, keratinocytes, fibroblasts, bronchial epithelial cells, and Reed-Sternberg cells. CCL17 promotes Th2 cell recruitment to sites of inflammation and induces platelet aggregation and degranulation.
  • Entrez Gene IDs:
    6361 (Human); 20295 (Mouse);
  • Aliases:
    ABCD-2; CC chemokine TARC; C-C motif chemokine 17; CCL17; chemokine (C-C motif) ligand 17; MGC138273; SCYA17MGC138271; small inducible cytokine subfamily A (Cys-Cys), member 17; Small-inducible cytokine A17; T cell-directed CC chemokine; TARC; TARCA-152E5.3; Thymus and activation-regulated chemokine
Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. 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.
  2. 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.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. 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.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. 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.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. 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.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. 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:

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.

10 Citations: Showing 1 - 10
Filter your results:

Species
Sample Type
  1. Forsythia suspensa Suppresses House Dust Mite Extract-Induced Atopic Dermatitis in NC/Nga Mice
    PLoS ONE, 2016;11(12):e0167687.
    Species: Human
    Sample Type: Cell Culture Supernates
  2. 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;10(2):e0116560.
    Species: Human
    Sample Type: Cell Culture Supernates
  3. 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;10(1):e0116775.
    Species: Human
    Sample Type: BALF
  4. 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;5(0):4092.
    Species: Human
    Sample Type: Cell Culture Supernates
  5. Marked and persistent eosinophilia in the absence of clinical manifestations.
    Authors: Chen Y, Khoury P, Ware J, Holland-Thomas N, Stoddard J, Gurprasad S, Waldner A, Klion A
    J Allergy Clin Immunol, 2014;133(4):1195-202.
    Species: Human
    Sample Type: Serum
  6. 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;186(7):3966-76.
    Species: Human
    Sample Type: Cell Culture Supernates
  7. Anemia in Hodgkin's lymphoma: the role of interleukin-6 and hepcidin.
    Authors: Hohaus S, Massini G, Giachelia M, Vannata B, Bozzoli V, Cuccaro A, D'Alo' F, Larocca LM, Raymakers RA, Swinkels DW, Voso MT, Leone G
    J. Clin. Oncol., 2010;28(15):2538-43.
    Species: Human
    Sample Type: Plasma
  8. Pathogenesis of cBFL in common with IPF? Correlation of IP-10/TARC ratio with histological patterns.
    Authors: Kishi M, Miyazaki Y, Jinta T, Furusawa H, Ohtani Y, Inase N, Yoshizawa Y
    Thorax, 2008;63(9):810-6.
    Species: Human
    Sample Type: Serum
  9. Respiratory syncytial virus synergizes with Th2 cytokines to induce optimal levels of TARC/CCL17.
    Authors: Monick MM, Powers LS, Hassan I, Groskreutz D, Yarovinsky TO, Barrett CW, Castilow EM, Tifrea D, Varga SM, Hunninghake GW
    J. Immunol., 2007;179(3):1648-58.
    Species: Human
    Sample Type: Cell Culture Supernates
  10. 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;110(13):4319-30.
    Species: Human
    Sample Type: Cell Culture Supernates
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