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    products/elisa/mouse-eotaxin-2-elisa-kit-ab206985.pdf

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Immunology Innate Immunity Chemokines Beta Chemokines (CC)
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RecombinantSimpleStep

Mouse Eotaxin-2 ELISA Kit (ab206985)

  • Datasheet
  • SDS
  • Protocol Booklet
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Other - Mouse Eotaxin-2 ELISA Kit (ab206985)
  • Example of the mouse Eotaxin-2 standard curve in Sample Diluent NS.
  • Linearity of dilution of mouse Eotaxin-2 in cell culture media and stimulated mouse lung supernatants.
  • Spiked linearity of dilution of mouse Eotaxin-2 in normal serum and normal plasma samples.
  • Native linearity of dilution of mouse Eotaxin-2 in normal serum and normal plasma samples.
  • Assay specificity is demonstrated on mouse lung supernatants.
  • Sandwich ELISA - Mouse Eotaxin-2 ELISA Kit (ab206985)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 6.23 pg/ml
  • Range: 19.53 pg/ml - 1250 pg/ml
  • Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Serum
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Mouse

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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, Cit 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%
    Cit plasma 95 89% - 102%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Goat, Cow, Pig
  • Product overview

    Mouse Eotaxin-2 ELISA Kit (ab206985) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Eotaxin-2 protein in cell culture supernatant, cit plasma, edta plasma, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse Eotaxin-2 with 6.23 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • 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.

  • 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 - HAMA Blocker (ab193969) 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 (ab193972) 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

    • Immunology
    • Innate Immunity
    • Chemokines
    • Beta Chemokines (CC)
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cytokines and cytokine receptors ELISA kits
  • 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

    • Entrez Gene: 56221 Mouse
    • SwissProt: Q9JKC0 Mouse

    Associated products

    • Alternative Versions

      • Mouse Eotaxin-2 Antibody Pair - BSA and Azide free (ab241690)

    Images

    • Other - Mouse Eotaxin-2 ELISA Kit (ab206985)
      Other - Mouse Eotaxin-2 ELISA Kit (ab206985)

      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.

       

    • Example of the mouse Eotaxin-2 standard curve in Sample Diluent NS.
      Example of the mouse Eotaxin-2 standard curve in Sample Diluent NS.

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

    • Linearity of dilution of mouse Eotaxin-2 in cell culture media and stimulated mouse lung supernatants.
      Linearity of dilution of mouse Eotaxin-2 in cell culture media and stimulated mouse lung supernatants.

      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).

    • Spiked linearity of dilution of mouse Eotaxin-2 in normal serum and normal plasma samples.
      Spiked linearity of dilution of mouse Eotaxin-2 in normal serum and normal plasma samples.

      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 linearity of dilution of mouse Eotaxin-2 in normal serum and normal plasma samples.
      Native linearity of dilution of mouse Eotaxin-2 in normal serum and normal plasma samples.

      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.

    • Assay specificity is demonstrated on mouse lung supernatants.
      Assay specificity is demonstrated on mouse lung supernatants.

      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.

    • Sandwich ELISA - Mouse Eotaxin-2 ELISA Kit (ab206985)
      Sandwich ELISA - Mouse Eotaxin-2 ELISA Kit (ab206985)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (1)

    Publishing research using ab206985? Please let us know so that we can cite the reference in this datasheet.

    ab206985 has been referenced in 1 publication.

    • Eberle JU  et al. Eosinophils Mediate Basophil-Dependent Allergic Skin Inflammation in Mice. J Invest Dermatol 139:1957-1965.e2 (2019). PubMed: 30910757

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