Overview

  • Product name

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

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 9 1.5%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 1.95%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    15.35 pg/ml
  • Range

    78.13 pg/ml - 5000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 106 102% - 110%
    Serum 102 97% - 105%
    Heparin Plasma 80 78% - 82%
    EDTA Plasma 102 94% - 107%
    Citrate Plasma 102 99% - 105%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    Human Eotaxin-2 SimpleStep ELISA® kit (ab179882) has been re-developed with new capture and detector antibodies. This new kit has the same name but a different product number (ab226898). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantifying Eotaxin-2 protein in human serum, plasma, and cell culture supernatant.


    Eotaxin-2 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Eotaxin-2 protein in human serum, plasma, and cell culture supernatant.


    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.


    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. 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

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BR 1 x 6ml
    10X Human Eotaxin-2 Capture Antibody 1 x 600µl
    10X Human Eotaxin-2 Detector Antibody 1 x 600µl
    Human 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

  • Function

    Chemotactic for resting T-lymphocytes, and eosinophils. Has lower chemotactic activity for neutrophils but none for monocytes and activated lymphocytes. Is a strong suppressor of colony formation by a multipotential hematopoietic progenitor cell line. Binds to CCR3.
  • Tissue specificity

    Activated monocytes and activated T lymphocytes.
  • Sequence similarities

    Belongs to the intercrine beta (chemokine CC) family.
  • Post-translational
    modifications

    N-glycosylated.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • C C motif chemokine 24
    • C-C motif chemokine 24
    • CCL24
    • CCL24_HUMAN
    • Chemokine CC Motif Ligand 24
    • CK beta 6
    • CK-beta-6
    • Ckb6
    • Eosinophil chemotactic protein 2
    • Eotaxin-2
    • MPIF 2
    • MPIF-2
    • MPIF2
    • Myeloid progenitor inhibitory factor 2
    • SCYA24
    • Small inducible cytokine A24
    • Small inducible cytokine subfamily A (Cys-Cys) member 24
    • Small-inducible cytokine A24
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab226898 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.

     

  •  Standard curve comparison between Human Eotaxin-2 SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows comparable sensitivity.

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

  • The concentrations of Eotaxin-2 were measured in duplicates, interpolated from the Eotaxin-2 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 3.12%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Eotaxin-2 concentration was determined to be 13,298 pg/mL in serum, 10,364 pg/mL in plasma (citrate), 9,354 pg/mL in plasma (EDTA), and 8,191 pg/mL in plasma (heparin).

  • PBMCs were cultured in RPMI media and either stimulated with 1.5% PHA or unstimulated for 48 hours. The concentrations of Eotaxin-2 were measured in duplicate and interpolated from the Eotaxin-2 standard curve and corrected for sample dilution. Undiluted samples are as follows: PBMC stimulated 3.12% and PBMC unstimulated 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Eotaxin-2 concentration was determined to be 107,776 pg/mL in stimulated PBMCs and 677 pg/mL in unstimulated PBMCs.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Eotaxin-2 concentration was determined to be 13,607 pg/mL with a range of 4,760 – 35,609 pg/mL.

Protocols

References

ab226898 has not yet been referenced specifically in any publications.

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