Overview

  • Product name
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    PBMC-PHA 5 4%
    Inter-assay
    Sample n Mean SD CV%
    PBMC-PHA 3 10%
  • Sample type
    Cell culture supernatant, Serum, Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    13 pg/ml
  • Range
    39.06 pg/ml - 2500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum = 85 83% - 85%
    Cell culture media = 104 96% - 111%
    Heparin Plasma = 88 84% - 91%
    EDTA Plasma = 80 79% - 81%
    Citrate Plasma = 98 82% - 107%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    MIP3a (CCL20, C-C motif chemokine 20) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the accurate quantitative measurement of MIP-3a protein in human cell culture supernatant, plasma and serum samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    CCL20 is a CC chemokine with a selective chemotactic activity for lymphocytes and dendritic cells. CCL20 inhibits proliferation of myeloid progenitors in colony formation assays. CCL20 may be involved in formation and function of the mucosal lymphoid tissues by attracting lymphocytes and dendritic cells towards epithelial cells. C-terminal processed forms of CCL20 have been shown to be equally chemotactically active for leukocytes. CCL20 is expressed predominantly in the liver, lymph nodes, appendix, peripheral blood lymphocytes, and fetal lung. CCL20 has low levels in thymus, prostate, testis, small intestine and colon. CCL20 is induced by bacterial lipopolysaccharides, TNF and IFNG/IFN-gamma. It is induced by phorbol myristate acetate (PMA) in U-937 cell line and bowes melanoma. It is repressed by IL10/interleukin-10. C-terminal processed forms of CCL20 which lack 1, 3 or 6 amino acids are produced by proteolytic cleavage after secretion from peripheral blood monocytes.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X MIP3a Capture Antibody 1 x 600µl
    10X MIP3a Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    MIP3a Human Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NBS 1 x 20ml
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Substrate 1 x 12ml
  • Research areas
  • Function
    Chemotactic factor that attracts lymphocytes and, slightly, neutrophils, but not monocytes. Inhibits proliferation of myeloid progenitors in colony formation assays. May be involved in formation and function of the mucosal lymphoid tissues by attracting lymphocytes and dendritic cells towards epithelial cells. C-terminal processed forms have been shown to be equally chemotactically active for leukocytes. Possesses antibacterial activity E.coli ATCC 25922 and S.aureus ATCC 29213.
  • Tissue specificity
    Expressed predominantly in the liver, lymph nodes, appendix, peripheral blood lymphocytes, and fetal lung. Low levels seen in thymus, prostate, testis, small intestine and colon.
  • Sequence similarities
    Belongs to the intercrine beta (chemokine CC) family.
  • Post-translational
    modifications
    C-terminal processed forms which lack 1, 3 or 6 amino acids are produced by proteolytic cleavage after secretion from peripheral blood monocytes.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Alternative names
    • Beta-chemokine exodus-1
    • C C motif chemokine ligand 20
    • C-C motif chemokine 20
    • CC chemokine LARC
    • Ccl20
    • CCL20(2-70)
    • CCL20_HUMAN
    • Chemokine (C C motif) ligand 20
    • Chemokine CC motif ligand 20
    • CKb4
    • Exodus
    • Exodus 1
    • LARC
    • Liver and activation-regulated chemokine
    • Macrophage inflammatory protein 3 alpha
    • MIP 3 alpha
    • MIP 3A
    • MIP-3-alpha
    • MIP-3a
    • MIP3A
    • SCYA20
    • Small inducible cytokine A20
    • Small inducible cytokine subfamily A (Cys Cys) member 20
    • Small-inducible cytokine A20
    • ST38
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab178015 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • PBMC were grown in the absence or presence of phytohemagglutinin (PHA) for 2 days. MIP3a concentrations were measured in 4X and 2X diluted and undiluted (neat) cell culture supernatants of the stimulated PBMC, unstimulated PBMC and media. Raw data values (mean +/ SD, n=5) are graphed. The dotted line represents zero sample background.

  • The concentrations of MIP3a were interpolated from data values shown in Figure 3 using MIP3a standard curve and corrected for sample dilution. Using 4X and 2X diluted samples, the mean MIP3a concentration in stimulated PBMC supernatants was determined to be 1,151 pg/mL. The undiluted sample of stimulated PBMC supernatant was not considered into the calculation because of poor recovery of MIP3a in 100% media (data not shown). MIP3a concentrations in unstimulated PBMC supernatants measured less than the MDD, 39 pg/mL.

Protocols

References

This product has been referenced in:
  • Menale C  et al. Bisphenol A effects on gene expression in adipocytes from children: association with metabolic disorders. J Mol Endocrinol 54:289-303 (2015). Read more (PubMed: 25878060) »

See 1 Publication for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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