Mouse CCL20/MIP-3 alpha DuoSet ELISA

Catalog # Availability Size / Price Qty
DY760
Ancillary Products Available
Mouse CCL20 / MIP-3 alpha ELISA Standard Curve
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Product Details
Procedure
Citations (11)
FAQs
Supplemental Products
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Mouse 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 mouse 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

Mouse CCL20 / MIP-3 alpha ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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 Mouse 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.

11 Citations: Showing 1 - 10
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  1. Interleukin 17A Promotes Lymphocytes Adhesion and Induces CCL2 and CXCL1 Release from Brain Endothelial Cells
    Authors: DW Wojkowska, P Szpakowski, A Glabinski
    Int J Mol Sci, 2017;18(5):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Klebsiella pneumoniae Siderophores Induce Inflammation, Bacterial Dissemination, and HIF-1? Stabilization during Pneumonia
    MBio, 2016;7(5):.
    Species: Mouse
    Sample Types: Tissue Homogenates
  3. Perinatal Activation of the Interleukin-33 Pathway Promotes Type 2 Immunity in the Developing Lung
    Authors: Ismé M de Kleer
    Immunity, 2016;45(6):1285-1298.
    Species: Mouse
    Sample Types: Tissue Homogenates
  4. Keratinocytes contribute intrinsically to psoriasis upon loss of Tnip1 function
    Proc Natl Acad Sci USA, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. 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: Mouse
    Sample Types: Cell Culture Supernates
  6. A TNF-alpha-CCL20-CCR6 axis regulates Nod1-induced B cell responses.
    Authors: Paradis M, Mindt B, Duerr C, Rojas O, Ng D, Boulianne B, McCarthy D, Yu M, Summers deLuca L, Ward L, Waldron J, Philpott D, Gommerman J, Fritz J
    J Immunol, 2014;192(6):2787-99.
    Species: Mouse
    Sample Types: Serum
  7. Nucleosomes contribute to increase mesangial cell chemokine expression during the development of lupus nephritis.
    Authors: Kanapathippillai P, Hedberg A, Fenton C, Fenton K
    Cytokine, 2013;62(2):244-52.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. Interleukin-1alpha controls allergic sensitization to inhaled house dust mite via the epithelial release of GM-CSF and IL-33.
    J. Exp. Med., 2012;209(8):1505-17.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  9. Toll-like receptor 4 mediates the response of epithelial and stromal cells to lipopolysaccharide in the endometrium.
    Authors: Sheldon IM, Roberts MH,
    PLoS ONE, 2010;5(9):e12906.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. A new mechanism for inhalational priming: IL-4 bypasses innate immune signals.
    Authors: Dittrich AM, Chen HC, Xu L, Ranney P, Connolly S, Yarovinsky TO, Bottomly HK
    J. Immunol., 2008;181(10):7307-15.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. Effect of oestradiol on PAMP-mediated CCL20/MIP-3 alpha production by mouse uterine epithelial cells in culture.
    Authors: Soboll G, Crane-Godreau MA, Lyimo MA, Wira CR
    Immunology, 2006;118(2):185-94.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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