Human CCL20/MIP-3 alpha Quantikine ELISA Kit

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DM3A00
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Human CCL20/MIP-3 alpha ELISA Calibrator Diluent RD6U Standard Curve
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Citations (42)
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Human 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 (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
Sensitivity
0.87 pg/mL
Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human 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 Human MIP-3 alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure MIP-3 alpha levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MIP-3 alpha and antibodies raised against the recombinant protein. Results obtained for naturally occurring human MIP-3 alpha showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural human 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

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 66.2 180 372 68.7 186 357
Standard Deviation 1.84 3.83 15.1 4.51 13 28.8
CV% 2.8 2.1 4.1 6.6 6.9 8.1

Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 55.9 161 323 72.5 200 386
Standard Deviation 1.62 4.28 9.1 6.05 17.9 35.3
CV% 2.9 2.7 2.8 8.3 8.9 9.1

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=8) 101 88-115
EDTA Plasma (n=5) 104 97-115
Heparin Plasma (n=5) 104 98-112
Serum (n=5) 103 94-112

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of MIP-3 alpha in various matrices were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CCL20/MIP-3 alpha ELISA Linearity

Data Examples

Human CCL20/MIP-3 alpha ELISA Calibrator Diluent RD6U Standard Curve

Human CCL20/MIP-3 alpha ELISA Calibrator Diluent RD6-21 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. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 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 Human 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.

42 Citations: Showing 1 - 10
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  1. Probiotic Lactobacillus and Bifidobacterium Strains Counteract Adherent-Invasive Escherichia coli (AIEC) Virulence and Hamper IL-23/Th17 Axis in Ulcerative Colitis, but Not in Crohn's Disease
    Authors: G Leccese, A Bibi, S Mazza, F Facciotti, F Caprioli, P Landini, M Paroni
    Cells, 2020;9(8):.
    Species: Human
    Sample Types: Whole Cells
  2. The EGFR-ERK/JNK-CCL20 Pathway in Scratched Keratinocytes May Underpin Koebnerization in Psoriasis Patients
    Authors: K Furue, T Ito, Y Tanaka, A Hashimoto-, M Takemura, M Murata, M Kido-Nakah, G Tsuji, T Nakahara, M Furue
    Int J Mol Sci, 2020;21(2):.
    Species: Human
    Sample Types: Cell Culture Supernatants
  3. Mucosal-associated invariant T cells redistribute to the peritoneal cavity during spontaneous bacterial peritonitis and contribute to peritoneal inflammation
    Authors: O Ibidapo-Ob, S Stengel, N Köse-Vogel, S Quickert, PA Reuken, M Busch, M Bauer, A Stallmach, T Bruns
    Cell Mol Gastroenterol Hepatol, 2020;0(0):.
    Species: Human
    Sample Types: Ascites Fluid
  4. Epigenetic downregulation of STAT6 increases HIF-1α expression via mTOR/S6K/S6, leading to enhanced hypoxic viability of glioma cells
    Authors: SJ Park, H Kim, SH Kim, EH Joe, I Jou
    Acta Neuropathol Commun, 2019;7(1):149.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Cyto/chemokine profile of in vitro scratched keratinocyte model: Implications of significant upregulation of CCL20, CXCL8 and IL36G in Koebner phenomenon
    Authors: K Furue, T Ito, Y Tanaka, A Yumine, A Hashimoto-, M Takemura, M Murata, K Yamamura, G Tsuji, M Furue
    J. Dermatol. Sci., 2019;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Staphylococcal Superantigens Stimulate Epithelial Cells through CD40 To Produce Chemokines
    Authors: PM Schlievert, MP Cahill, BS Hostager, AJ Brosnahan, AJ Klingelhut, FA Gourronc, GA Bishop, DYM Leung
    MBio, 2019;10(2):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Th17-associated cytokines multiplex testing indicates the potential of macrophage inflammatory protein-3 alpha in the diagnosis of biliary atresia
    Authors: P Chen, Z Zhong, H Jiang, H Chen, J Lyu, L Zhou
    Cytokine, 2019;116(0):21-26.
    Species: Human
    Sample Types: Serum
  8. Effects of modified FOLFOX-6 chemotherapy on cellular immune function in patients with gastric cancer
    Authors: L Wang, D Zhou, H Ren, Y Chen
    Oncol Lett, 2018;15(6):8635-8640.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Fusobacterium nucleatum subspecies animalis influences pro-inflammatory cytokine expression and monocyte activation in human colorectal tumors
    Authors: X Ye, R Wang, R Bhattachar, D Boulbes, F Fan, L Xia, H Adoni, NJ Ajami, MC Wong, DP Smith, JF Petrosino, S Venable, W Qiao, V Baladanday, DM Maru, LM Ellis
    Cancer Prev Res (Phila), 2017;0(0):.
    Species: Human
    Sample Types: Tissue Homogenates
  10. CCL20 secreted from IgA1-stimulated human mesangial cells recruits inflammatory Th17 cells in IgA nephropathy
    Authors: G Lu, X Zhang, L Shen, Q Qiao, Y Li, J Sun, J Zhang
    PLoS ONE, 2017;12(5):e0178352.
    Species: Human
    Sample Types: Serum
  11. CCL20 mediates RANK/RANKL-induced epithelial-mesenchymal transition in endometrial cancer cells
    Oncotarget, 2016;7(18):25328-39.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Impairment of CCR6+ and CXCR3+ Th Cell Migration in HIV-1 Infection Is Rescued by Modulating Actin Polymerization
    Authors: Valentina Cecchinato
    J. Immunol, 2016;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. CCL20-CCR6 Cytokine Network Facilitate Treg Activity in Advanced Grades and Metastatic Variants of Hepatocellular Carcinoma.
    Authors: Li W, Liu H
    Scand J Immunol, 2015;83(1):33-7.
    Species: Human
    Sample Types: Tissue Homogenates
  14. Autophagy facilitates TLR4- and TLR3-triggered migration and invasion of lung cancer cells through the promotion of TRAF6 ubiquitination.
    Authors: Zhan Z, Xie X, Cao H, Zhou X, Zhang X, Fan H, Liu Z
    Autophagy, 2014;10(2):257-68.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Selective impact of HIV disease progression on the innate immune system in the human female reproductive tract.
    Authors: Lahey T, Ghosh M, Fahey JV, Shen Z, Mukura LR, Song Y, Cu-Uvin S, Mayer KH, Wright PF, Kappes JC, Ochsenbauer C, Wira CR
    PLoS ONE, 2012;7(6):e38100.
    Species: Human
    Sample Types: Vaginal Lavage Fluid
  16. A randomized trial to assess anti-HIV activity in female genital tract secretions and soluble mucosal immunity following application of 1% tenofovir gel.
    Authors: Keller MJ, Madan RP, Torres NM
    PLoS ONE, 2011;6(1):e16475.
    Species: Human
    Sample Types: Vaginal Lavage Fluid
  17. IL-22, but not IL-17, dominant environment in cutaneous T-cell lymphoma.
    Authors: Miyagaki T, Sugaya M, Suga H, Kamata M, Ohmatsu H, Fujita H, Asano Y, Tada Y, Kadono T, Sato S
    Clin. Cancer Res., 2011;17(24):7529-38.
    Species: Human
    Sample Types: Serum
  18. Opposing roles of IL-17A and IL-25 in the regulation of TSLP production in human nasal epithelial cells.
    Authors: Xu G, Zhang L, Wang DY, Xu R, Liu Z, Han DM, Wang XD, Zuo KJ, Li HB
    Allergy, 2010;65(5):581-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Activation of the aryl hydrocarbon receptor reveals distinct requirements for IL-22 and IL-17 production by human T helper cells.
    Authors: Ramirez JM, Brembilla NC, Sorg O, Chicheportiche R, Matthes T, Dayer JM, Saurat JH, Roosnek E, Chizzolini C
    Eur. J. Immunol., 2010;40(9):2450-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Characterization of circulating dendritic cells in melanoma: role of CCR6 in plasmacytoid dendritic cell recruitment to the tumor.
    Authors: Charles J, Di Domizio J, Salameire D, Bendriss-Vermare N, Aspord C, Muhammad R, Lefebvre C, Plumas J, Leccia MT, Chaperot L
    J. Invest. Dermatol., 2010;130(6):1646-56.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. HLA-DR+ leukocytes acquire CD1 antigens in embryonic and fetal human skin and contain functional antigen-presenting cells.
    Authors: Schuster C, Vaculik C, Fiala C, Meindl S, Brandt O, Imhof M, Stingl G, Eppel W, Elbe-Burger A
    J. Exp. Med., 2009;206(1):169-81.
    Species: Human
    Sample Types: Cell Culture Supernates
  22. Intratumoral induction of CD103 triggers tumor-specific CTL function and CCR5-dependent T-cell retention.
    Authors: Franciszkiewicz K, Le Floc'h A, Jalil A, Vigant F, Robert T, Vergnon I, Mackiewicz A, Benihoud K, Validire P, Chouaib S, Combadiere C, Mami-Chouaib F
    Cancer Res., 2009;69(15):6249-55.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Prostaglandin E2 regulates Th17 cell differentiation and function through cyclic AMP and EP2/EP4 receptor signaling.
    Authors: Boniface K, Bak-Jensen KS, Li Y, Blumenschein WM, McGeachy MJ, McClanahan TK, McKenzie BS, Kastelein RA, Cua DJ, de Waal Malefyt R
    J. Exp. Med., 2009;206(3):535-48.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. The imbalance in serum concentration of Th-1- and Th-2-derived chemokines as one of the factors involved in pathogenesis of atopic dermatitis.
    Authors: Narbutt J, Lesiak A, Sysa-Jedrzeiowska A, Zakrzewski M, Bogaczewicz J, Stelmach I, Kuna P
    Mediators Inflamm., 2009;2009(0):269541.
    Species: Human
    Sample Types: Serum
  25. Genome-wide comparison between IL-17A- and IL-17F-induced effects in human rheumatoid arthritis synoviocytes.
    Authors: Zrioual S, Ecochard R, Tournadre A, Lenief V, Cazalis MA, Miossec P
    J. Immunol., 2009;182(0):3112-20.
    Species: Human
    Sample Types: Cell Culture Supernates
  26. 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: Human
    Sample Types: Cell Culture Supernates
  27. IL-17RA and IL-17RC receptors are essential for IL-17A-induced ELR+ CXC chemokine expression in synoviocytes and are overexpressed in rheumatoid blood.
    Authors: Zrioual S, Toh ML, Tournadre A, Zhou Y, Cazalis MA, Pachot A, Miossec V, Miossec P
    J. Immunol., 2008;180(1):655-63.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. Macrophage inflammatory protein-3alpha is a novel serum marker for nasopharyngeal carcinoma detection and prediction of treatment outcomes.
    Authors: Chang KP, Hao SP, Chang JH, Wu CC, Tsang NM, Lee YS, Hsu CL, Ueng SH, Liu SC, Liu YL, Wei PC, Liang Y, Chang YS, Yu JS
    Clin. Cancer Res., 2008;14(21):6979-87.
    Species: Human
    Sample Types: Serum
  29. Urinary fractalkine is a marker of acute rejection.
    Authors: Peng W, Chen J, Jiang Y, Wu J, Shou Z, He Q, Wang Y, Chen Y, Wang H
    Kidney Int., 2008;74(11):1454-60.
    Species: Human
    Sample Types: Urine
  30. Expression of the Epstein-Barr virus-encoded Epstein-Barr virus nuclear antigen 1 in Hodgkin's lymphoma cells mediates Up-regulation of CCL20 and the migration of regulatory T cells.
    Authors: Baumforth KR, Birgersdotter A, Reynolds GM, Wei W, Kapatai G, Flavell JR, Kalk E, Piper K, Lee S, Machado L, Hadley K, Sundblad A, Sjoberg J, Bjorkholm M, Porwit AA, Yap LF, Teo S, Grundy RG, Young LS, Ernberg I, Woodman CB, Murray PG
    Am. J. Pathol., 2008;173(1):195-204.
    Species: Human
    Sample Types: Cell Culture Supernates
  31. Chemokine production and chemokine receptor expression by human glioma cells: role of CXCL10 in tumour cell proliferation.
    Authors: Maru SV, Holloway KA, Flynn G, Lancashire CL, Loughlin AJ, Male DK, Romero IA
    J. Neuroimmunol., 2008;199(1):35-45.
    Species: Human
    Sample Types: Cell Culture Supernates
  32. Evidence for a pathogenetic role of interleukin-18 in cutaneous lupus erythematosus.
    Authors: Wang D, Drenker M, Eiz-Vesper B, Werfel T, Wittmann M
    Arthritis Rheum., 2008;58(10):3205-15.
    Species: Human
    Sample Types: Cell Culture Supernates
  33. Accumulation of dendritic cells and increased CCL20 levels in the airways of patients with chronic obstructive pulmonary disease.
    Authors: Demedts IK, Bracke KR, Van Pottelberge G, Testelmans D, Verleden GM, Vermassen FE, Joos GF, Brusselle GG
    Am. J. Respir. Crit. Care Med., 2007;175(10):998-1005.
    Species: Human
    Sample Types: Sputum
  34. Airway dendritic cell phenotypes in inflammatory diseases of the human lung.
    Authors: Lommatzsch M, Bratke K, Bier A, Julius P, Kuepper M, Luttmann W, Virchow JC
    Eur. Respir. J., 2007;30(5):878-86.
    Species: Human
    Sample Types: BALF
  35. Specificity of immunomodulator secretion in urinary samples in response to infection by alpha-hemolysin and CNF1 bearing uropathogenic Escherichia coli.
    Authors: Real JM, Munro P, Buisson-Touati C, Lemichez E, Boquet P, Landraud L
    Cytokine, 2007;37(1):22-5.
    Species: Human
    Sample Types: Urine
  36. Glycerol monolaurate inhibits the effects of Gram-positive select agents on eukaryotic cells.
    Authors: Peterson ML, Schlievert PM
    Biochemistry, 2006;45(7):2387-97.
    Species: Human
    Sample Types: Cell Culture Supernates
  37. Hypoxia modifies the transcriptome of primary human monocytes: modulation of novel immune-related genes and identification of CC-chemokine ligand 20 as a new hypoxia-inducible gene.
    Authors: Bosco MC, Puppo M, Santangelo C, Anfosso L, Pfeffer U, Fardin P, Battaglia F, Varesio L
    J. Immunol., 2006;177(3):1941-55.
    Species: Human
    Sample Types: Cell Culture Supernates
  38. Expression of CCL20 and granulocyte-macrophage colony-stimulating factor, but not Flt3-L, from modified vaccinia virus ankara enhances antiviral cellular and humoral immune responses.
    Authors: Chavan R, Marfatia KA, An IC, Garber DA, Feinberg MB
    J. Virol., 2006;80(15):7676-87.
    Species: Human
    Sample Types: Cell Culture Supernates
  39. 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: Human
    Sample Types: Cell Culture Supernates
  40. Trichomonas vaginalis lipophosphoglycan triggers a selective upregulation of cytokines by human female reproductive tract epithelial cells.
    Authors: Fichorova RN, Trifonova RT, Gilbert RO, Costello CE, Hayes GR, Lucas JJ, Singh BN
    Infect. Immun., 2006;74(10):5773-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  41. Role for cysteinyl leukotrienes in allergen-induced change in circulating dendritic cell number in asthma.
    Authors: Parameswaran K, Liang H, Fanat A, Watson R, Snider DP, O&amp;amp;apos;Byrne PM
    J. Allergy Clin. Immunol., 2004;114(1):73-9.
    Species: Human
    Sample Types: Sputum
  42. Plasticity of airway epithelial cell transcriptome in response to flagellin.
    Authors: Clark J, Kim K, Basom R, Gharib S
    PLoS ONE, 0;10(2):e0115486.
    Species: Human
    Sample Types: Cell Culture Supernates

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Human CCL20/MIP-3 alpha Quantikine ELISA Kit
By Anonymous on 01/20/2017
Application: Sample Tested: Cell Culture Supernates

This kit is a very nice tool in determining the level of MIP-3alpha in the cell culture spent media.


Human CCL20/MIP-3 alpha Quantikine ELISA Kit
By Anonymous on 01/03/2017
Application: Sample Tested: Stimulated Human Bronchial Epithelial Cell Line (BEAS-2B)

This kit is an excellent tool in determining the level of MIP-3a in the culture media.