Mouse CCL20/MIP-3 alpha Quantikine ELISA Kit

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MCC200
Mouse CCL20/MIP-3 alpha ELISA Standard Curve
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Citations (16)
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Mouse CCL20/MIP-3 alpha Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), Heparin Plasma (50 uL)
Sensitivity
0.49 pg/mL
Assay Range
3.9 - 250 pg/mL (Cell Culture Supernates, Serum, Heparin Plasma)
Specificity
Natural and recombinant mouse MIP-3 alpha
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Mouse MIP-3 alpha Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse MIP-3 alpha in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse MIP-3 alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse MIP-3 alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse MIP-3 alpha.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 23 23 23
Mean (pg/mL) 10 43 118 11 40 112
Standard Deviation 0.6 1.7 5.9 1 3.6 7.5
CV% 6 4 5 9.1 9 6.7

Recovery

The recovery of mouse MIP-3 alpha spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 100 96-106
Heparin Plasma (n=4) 98 85-115
Serum (n=4) 106 95-119

Linearity

To assess the linearity of the assay, samples spiked with recombinant mouse MIP-3 alpha in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse CCL20/MIP-3 alpha ELISA Linearity

Data Examples

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.
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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Mouse CCL20/MIP-3 alpha Quantikine ELISA Kit

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.

16 Citations: Showing 1 - 10
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  1. Tumor suppressor microRNA-204-5p regulates growth, metastasis, and immune microenvironment remodeling in breast cancer
    Authors: BS Hong, HS Ryu, N Kim, J Kim, E Lee, H Moon, KH Kim, MS Jin, NH Kwon, S Kim, D Kim, DH Chung, K Jeong, K Kim, KY Kim, HB Lee, W Han, J Yun, JI Kim, DY Noh, HG Moon
    Cancer Res., 2019;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. CCL20-CCR6 axis modulated traumatic brain injury-induced visual pathologies
    Authors: M Das, X Tang, JY Han, K Mayilsamy, E Foran, MR Biswal, R Tzekov, SS Mohapatra, S Mohapatra
    J Neuroinflammation, 2019;16(1):115.
    Species: Mouse
    Sample Types: Tissue Homogenates
  3. Obesity exacerbates colitis-associated cancer via IL-6-regulated macrophage polarisation and CCL-20/CCR-6-mediated lymphocyte recruitment
    Authors: CM Wunderlich, PJ Ackermann, AL Ostermann, P Adams-Quac, MC Vogt, ML Tran, A Nikolajev, A Waisman, C Garbers, S Theurich, J Mauer, N Hövelmeyer, FT Wunderlich
    Nat Commun, 2018;9(1):1646.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Lung Epithelial Cells Coordinate Innate Lymphocytes and Immunity against Pulmonary Fungal Infection
    Authors: N Hernández-, DL Wiesner, JS Fites, AJ McDermott, T Warner, M Wüthrich, BS Klein
    Cell Host Microbe, 2018;0(0):.
    Species: Mouse
    Sample Types: BALF
  5. Prion infection of mouse brain reveals multiple new upregulated genes involved in neuroinflammation or signal transduction.
    Authors: Carroll J, Striebel J, Race B, Phillips K, Chesebro B
    J Virol, 2015;89(4):2388-404.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. Leukocyte attraction by CCL20 and its receptor CCR6 in humans and mice with pneumococcal meningitis.
    Authors: Klein M, Brouwer M, Angele B, Geldhoff M, Marquez G, Varona R, Hacker G, Schmetzer H, Hacker H, Hammerschmidt S, van der Ende A, Pfister H, van de Beek D, Koedel U
    PLoS ONE, 2014;9(4):e93057.
    Species: Mouse
    Sample Types: Tissue Homogenates
  7. Potential roles of CCR5(+) CCR6(+) dendritic cells induced by nasal ovalbumin plus Flt3 ligand expressing adenovirus for mucosal IgA responses.
    Authors: Fukuyama Y, Tokuhara D, Sekine S, Aso K, Kataoka K, Davydova J, Yamamoto M, Gilbert R, Tokuhara Y, Fujihashi K, Kunisawa J, Yuki Y, Kiyono H, McGhee J, Fujihashi K
    PLoS ONE, 2013;8(4):e60453.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  8. Monocyte-derived dendritic cell recruitment and allergic T(H)2 responses after exposure to diesel particles are CCR2 dependent.
    Authors: Provoost S, Maes T, Joos GF, Tournoy KG
    J. Allergy Clin. Immunol., 2012;129(2):483-91.
    Species: Mouse
    Sample Types: BALF
  9. The Role of ChemR23 in the Induction and Resolution of Cigarette Smoke-Induced Inflammation.
    Authors: Demoor T, Bracke KR, Dupont LL
    J. Immunol., 2011;186(9):5457-67.
    Species: Mouse
    Sample Types: BALF
  10. CCL20, gammadelta T cells, and IL-22 in corneal epithelial healing.
    Authors: Li Z, Burns AR, Miller SB, Smith CW
    FASEB J., 2011;25(8):2659-68.
    Species: Mouse
    Sample Types: Tissue Homogenates
  11. NOD1 contributes to mouse host defense against Helicobacter pylori via induction of type I IFN and activation of the ISGF3 signaling pathway.
    Authors: Watanabe T, Asano N, Fichtner-Feigl S, Gorelick PL, Tsuji Y, Matsumoto Y, Chiba T, Fuss IJ, Kitani A, Strober W
    J. Clin. Invest., 2010;120(5):1645-62.
    Species: Mouse
    Sample Types: Tissue Homogenates
  12. Th17 Cytokines Stimulate CCL20 Expression in Keratinocytes In Vitro and In Vivo: Implications for Psoriasis Pathogenesis.
    Authors: Harper EG, Guo C, Rizzo H, Lillis JV, Kurtz SE, Skorcheva I, Purdy D, Fitch E, Iordanov M, Blauvelt A
    J. Invest. Dermatol., 2009;129(0):2175.
    Species: Mouse
    Sample Types: Tissue Homogenates
  13. Mouse models of rhinovirus-induced disease and exacerbation of allergic airway inflammation.
    Authors: Bartlett NW, Walton RP, Edwards MR, Aniscenko J, Caramori G, Zhu J, Glanville N, Choy KJ, Jourdan P, Burnet J, Tuthill TJ, Pedrick MS, Hurle MJ, Plumpton C, Sharp NA, Bussell JN, Swallow DM, Schwarze J, Guy B, Almond JW, Jeffery PK, Lloyd CM, Papi A, Killington RA, Rowlands DJ, Blair ED, Clarke NJ, Johnston SL
    Nat. Med., 2008;14(2):199-204.
    Species: Mouse
    Sample Types: BALF
  14. Tyrosine phosphatase SHP-1 in oxidative stress and development of allergic airway inflammation.
    Authors: Cho YS, Oh SY, Zhu Z
    Am. J. Respir. Cell Mol. Biol., 2008;39(4):412-9.
    Species: Mouse
    Sample Types: BALF
  15. Chemokine receptor CCR2 but not CCR5 or CCR6 mediates the increase in pulmonary dendritic cells during allergic airway inflammation.
    Authors: Robays LJ, Maes T, Lebecque S, Lira SA, Kuziel WA, Brusselle GG, Joos GF, Vermaelen KV
    J. Immunol., 2007;178(8):5305-11.
    Species: Mouse
    Sample Types: BALF
  16. An inflammation-induced mechanism for leukocyte transmigration across lymphatic vessel endothelium.
    Authors: Johnson LA, Clasper S, Holt AP, Lalor PF, Baban D, Jackson DG
    J. Exp. Med., 2006;203(12):2763-77.
    Species: Mouse
    Sample Types: Cell Culture Supernates

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