Overview

  • Product name

    Mouse Eotaxin-2 ELISA Kit
    See all Eotaxin 2 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 8 3.42%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 4.66%
  • Sample type

    Cell culture supernatant, Serum, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    6.23 pg/ml
  • Range

    19.53 pg/ml - 1250 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 100 98% - 101%
    Cell culture media 94 90% - 99%
    EDTA Plasma 98 94% - 105%
    Citrate Plasma 95 89% - 102%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rat, Rabbit, Goat, Guinea pig, Hamster, Cow, Dog, Human, Pig
  • Product overview

    Abcam’s Eotaxin-2 (CCL24) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Eotaxin-2  in mouse serum, plasma, and cell culture supernatants.

    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

    Eotaxin-2, also known as CCL24, MPIF-2 or Ckβ-6, is a C-C chemokine originally identified in activated monocytes. Eotaxin-2 is expressed in activated T lymphocytes, GM-CSF treated macrophages and dermal fibroblasts. Mouse eotaxin-2 is a highly glycosylated protein which requires maturation by removal of a 26 aminoacid signal peptide prior to secretion. Secretion of Eotaxin-2 is known to recruit and activate CCR3-bearing cells particularly eosinophils as well as basophils. Activation of eosinophils generates reactive oxygen species, lipid mediators of inflammation and degranulation of toxic granule proteins. Chemotaxis and activation of basophils, previously primed with IL-3, lead to release of histamine and leukotriene C4.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse Eotaxin-2 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPI 1 x 6ml
    Mouse Eotaxin-2 Capture Antibody (lyophilized) 1 vial
    Mouse Eotaxin-2 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Relevance

    Eotaxin 2 is a member of the CC chemokine family, based on the presence of the CC motif and homology with other known CC chemokines. Eotaxin 2 cDNA encodes a 119 amino acid residue precursor protein with a 26 amino acid residue signal peptide that is cleaved to generate a mature protein predicted to contain 93 amino acid residues with an N glycosylation site. Mature human eotaxin 2 has a predicted molecular mass of approximately 10.6 kDa. Compared to other CC chemokines, eotaxin 2 exhibits 40 %, 42 %, and 39 % amino acid identity to MCP3, MIP1 alpha, and eotaxin, respectively. Human CC chemokine eotaxin 2 maps to chromosome 7q11.23. Both eotaxin and eotaxin 2 activate and attract eosinophils and basophils. A receptor for human eotaxin has been identified and found to be the third numbered receptor in the C-C chemokine subfamily of receptors (CCR3. On eosinophils, the effects of eotaxin 2 is inhibited by an CCR3 antibody and cross-desensitized by eotaxin and MCP4, suggesting that all three CC chemokines act through CCR3. Eotaxin 2 mRNA is weakly expressed in activated monocytes and T lymphocytes. Recombinant eotaxin 2 induces chemotaxis of eosinophils, basophils, and resting T lymphocytes but not monocytes and activated T lymphocytes. Eotaxin 2 inhibits colony formation in myleloid progenitor cells.
  • Cellular localization

    Secreted
  • Alternative names

    • C C motif chemokine 24
    • CCL24
    • Chemokine (C C motif) ligand 24
    • Chemokine CC Motif Ligand 2
    • CK beta 6
    • Ckb 6
    • Ckb6
    • Eosinophil chemotactic protein 2
    • Eotaxin 2 like protein
    • MPIF 2
    • MPIF2
    • Myeloid progenitor inhibitory factor 2
    • OTTHUMP00000210225
    • SCYA24
    • Small inducible cytokine A24
    • Small inducible cytokine subfamily A (Cys Cys) member 24
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab206985 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

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

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

  • Recombinant mouse Eotaxin-2 was spiked into 1:4 cell culture media and diluted in a 2-fold dilution series in Sample Diluent NS. Native mouse Eotaxin-2 was measured in 1:20 stimulated mouse lung supernatants which was further diluted in a 2-fold dilution series in Sample Diluent NS. The concentrations of mouse Eotaxin-2 were measured in duplicate and interpolated from the mouse Eotaxin-2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • Recombinant mouse Eotaxin-2 was spiked into neat serum, neat citrate plasma, and neat EDTA plasma diluted in a 2-fold dilution series in Sample Diluent NS. The concentrations of mouse Eotaxin-2 were measured in duplicate and interpolated from the mouse Eotaxin-2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are graphed (mean +/- SD).

  • Native mouse Eotaxin-2 was measured in neat normal serum, neat normal citrate plasma, and neat normal EDTA plasma diluted in a 2-fold dilution series in Sample Diluent NS. The concentrations of mouse Eotaxin-2 were measured in duplicate and interpolated from the mouse Eotaxin 2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are graphed (mean +/- SD). For reference, 8X or more diluted normal serum and normal citrate plasma samples interpolate below the standard curve. 16X or more diluted Normal EDTA plasma samples interpolate below the standard curve.

  • Mouse lung supernatants were cultured in cell culture media with 10% fetal bovine serum for 3 days in the absence (unstimulated) and presence (stimulated) of 10 ng/mL of human IL-4. Both unstimulated and stimulated mouse lung supernatants were measured in duplicate at a 1:20 dilution. The Raw O.D. values for each sample are graphed, with the background O.D. shown as the dashed line. The stimulated mouse lung supernatants diluted 1:20 measured at 1,207 pg/mL, whereas the unstimulated mouse lung supernatants diluted 1:20 measured below the limit of detection of the assay.

Protocols

References

ab206985 has not yet been referenced specifically in any publications.

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