Human CCL20/MIP-3 alpha DuoSet ELISA

Catalog # Availability Size / Price Qty
DY360
Ancillary Products Available
Human CCL20 / MIP-3 alpha ELISA Standard Curve
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Product Details
Procedure
Citations (19)
FAQs
Supplemental Products
Reviews (1)

Human CCL20/MIP-3 alpha DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CCL20/MIP-3alpha. 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)

Data Example

Human CCL20 / MIP-3 alpha ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CCL20/MIP-3 alpha

CCL20, also known as MIP-3 alpha, is a chemokine that plays an important role in dendritic cell trafficking and recruitment and activation of T cells. It is upregulated in monocytes, T cells, endothelial cells, epithelial cells, and fibroblasts following inflammatory stimulation. CCL20 signals through CCR6 which is expressed on memory T cells, B cells and bone marrow-derived dendritic cells.

Entrez Gene IDs:
6364 (Human); 20297 (Mouse); 29538 (Rat)
Alternate Names:
beta chemokine exodus-1; Beta-chemokine exodus-1; CC chemokine LARC; C-C motif chemokine 20; CCL20; chemokine (C-C motif) ligand 20; CKb4; exodus-1; LARC; LARCLiver and activation-regulated chemokine; MIP3 alpha; MIP-3 alpha; MIP-3a; MIP-3-alpha; MIP3AMacrophage inflammatory protein 3 alpha; SCYA20Small-inducible cytokine A20; small inducible cytokine subfamily A (Cys-Cys), member 20; ST38

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 for Human CCL20/MIP-3 alpha 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.

19 Citations: Showing 1 - 10
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  1. SIRT1/FoxO3 axis alteration leads to aberrant immune responses in bronchial epithelial cells
    Authors: S Di Vincenz, IH Heijink, JA Noordhoek, C Cipollina, L Siena, A Bruno, M Ferraro, DS Postma, M Gjomarkaj, E Pace
    J. Cell. Mol. Med., 2018;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Distinctive expression of T cell guiding molecules in human autoimmune lymph node stromal cells upon TLR3 triggering
    Authors: JS Hähnlein, TH Ramwadhdoe, JF Semmelink, IY Choi, FH Berger, M Maas, DM Gerlag, PP Tak, TBH Geijtenbee, LGM van Baarse
    Sci Rep, 2018;8(1):1736.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Rheumatoid arthritis bone marrow environment supports Th17 response
    Authors: E Kuca-Warna, W Kurowska, M Prochorec-, A Radzikowsk, T Burakowski, U Skalska, M Massalska, M Pleba?czyk, B Ma?dyk-Now, I S?owi?ska, R Gasik, W Ma?li?ski
    Arthritis Res. Ther., 2017;19(1):274.
    Species: Human
    Sample Types: Plasma
  4. IL-27 Modulates Chemokine Production in TNF-? -Stimulated Human Oral Epithelial Cells
    Authors: Y Hosokawa, I Hosokawa, K Ozaki, T Matsuo
    Cell. Physiol. Biochem., 2017;43(3):1198-1206.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Human Lymph Node-Derived Fibroblastic and Double-Negative Reticular Cells Alter Their Chemokines and Cytokines Expression Profile Following Inflammatory Stimuli
    Authors: P Severino, DT Palomino, H Alvarenga, CB Almeida, DC Pasqualim, A Cury, PR Salvalaggi, AL De Vasconc, MC Andrade, T Aloia, S Bromberg, LV Rizzo, FA Rocha, LC Marti
    Front Immunol, 2017;8(0):141.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Transforming Growth Factor-beta and Interleukin-1beta Signaling Pathways Converge on the Chemokine CCL20 Promoter.
    Authors: Brand O, Somanath S, Moermans C, Yanagisawa H, Hashimoto M, Cambier S, Markovics J, Bondesson A, Hill A, Jablons D, Wolters P, Lou J, Marks J, Baron J, Nishimura S
    J Biol Chem, 2015;290(23):14717-28.
    Species: Human
    Sample Types: Tissue Homogenates
  7. Bacterial siderophores that evade or overwhelm lipocalin 2 induce hypoxia inducible factor 1alpha and proinflammatory cytokine secretion in cultured respiratory epithelial cells.
    Authors: Holden V, Lenio S, Kuick R, Ramakrishnan S, Shah Y, Bachman M
    Infect Immun, 2014;82(9):3826-36.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. 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;8(12):e82867.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Human Papillomavirus Type 8 Interferes with a Novel C/EBPβ-Mediated Mechanism of Keratinocyte CCL20 Chemokine Expression and Langerhans Cell Migration.
    Authors: Sperling T, Oldak M, Walch-Ruckheim B, Wickenhauser C, Doorbar J, Pfister H, Malejczyk M, Majewski S, Keates AC, Smola S
    PLoS Pathog., 2012;8(7):e1002833.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. 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 Types: Cell Culture Supernates
  11. Candida albicans yeast and hyphae are discriminated by MAPK signaling in vaginal epithelial cells.
    Authors: Moyes DL, Murciano C, Runglall M, Islam A, Thavaraj S, Naglik JR
    PLoS ONE, 2011;6(11):e26580.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. CCL20/CCR6 feedback exaggerates epidermal growth factor receptor-dependent MUC5AC mucin production in human airway epithelial (NCI-H292) cells.
    Authors: Kim S, Lewis C, Nadel JA
    J. Immunol., 2011;186(6):3392-400.
    Species: Human
    Sample Types: Cell Lysates
  13. Interleukin-17 and interferon-gamma are produced concomitantly by human coronary artery-infiltrating T cells and act synergistically on vascular smooth muscle cells.
    Authors: Eid RE, Rao DA, Zhou J, Lo SF, Ranjbaran H, Gallo A, Sokol SI, Pfau S, Pober JS, Tellides G
    Circulation, 2009;119(10):1424-32.
    Species: Human
    Sample Types: Plasma
  14. IL-17 and IL-22 mediate IL-20 subfamily cytokine production in cultured keratinocytes via increased IL-22 receptor expression.
    Authors: Tohyama M, Hanakawa Y, Shirakata Y, Dai X, Yang L, Hirakawa S, Tokumaru S, Okazaki H, Sayama K, Hashimoto K
    Eur. J. Immunol., 2009;39(10):2779-88.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Lactoferrin, a major defense protein of innate immunity, is a novel maturation factor for human dendritic cells.
    Authors: Spadaro M, Caorsi C, Ceruti P, Varadhachary A, Forni G, Pericle F, Giovarelli M
    FASEB J., 2008;22(8):2747-57.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Lipopolysaccharide-squamous cell carcinoma-monocyte interactions induce cancer-supporting factors leading to rapid STAT3 activation.
    Authors: Kurago ZB, Lam-ubol A, Stetsenko A
    Head Neck Pathol, 2008;2(1):1-12.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. The role of a nuclear protein, histone H1, on signalling pathways for the maturation of dendritic cells.
    Authors: Hsu LW, Chen CL, Nakano T, Lai CY, Chiang KC, Lin YC, Kao YH, Chen SH, Goto T, Sung WC, Yang CH, Cheng YF, Jawan B, Chiu KW, Goto S
    Clin. Exp. Immunol., 2008;152(3):576-84.
    Species: Human
    Sample Types: BALF
  18. Multiple NF-kappaB and IFN regulatory factor family transcription factors regulate CCL19 gene expression in human monocyte-derived dendritic cells.
    Authors: Pietila TE, Veckman V, Lehtonen A, Lin R, Hiscott J, Julkunen I
    J. Immunol., 2007;178(1):253-61.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. The effects of IL-20 subfamily cytokines on reconstituted human epidermis suggest potential roles in cutaneous innate defense and pathogenic adaptive immunity in psoriasis.
    Authors: Sa SM, Valdez PA, Wu J
    J. Immunol., 2007;178(4):2229-40.
    Species: Human
    Sample Types: Cell Culture Supernates

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Human CCL20/MIP-3 alpha DuoSet ELISA
By Anonymous on 08/23/2016
Sample Tested: Purified Standard