Overview

  • Product name

    Mouse MCP2 ELISA Kit (CCL8)
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 8 3.98%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 2.99%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    1 pg/ml
  • Range

    2.34 pg/ml - 150 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 101 97% - 104%
    Cell culture media 84 82% - 86%
    Heparin Plasma 99 90% - 103%
    EDTA Plasma 107 105% - 109%
    Citrate Plasma 108 108% - 110%

  • 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

    MCP2 (CCL8) in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of MCP2 protein in mouse serum, plasma, cell culture supernatants, and tissue extracts.


    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

    MCP2 is a chemotactic factor that attracts monocytes. Chemokines and their receptors, in conjunction with adhesion molecules and their ligands, provide cues to enable tissue-specific homing of T cells in the steady state and during inflammation. Mouse MCP specifically induces migration of a population of recently activated, highly differentiated TH2 cells enriched in interleukin 5 (IL-5) and the IL-25 receptor (IL-25R), cytokines implicated in eosinophilic inflammation; as well as tumor necrosis factor (TNF) and OX40, molecules associated with inflammatory TH2 cells. Using a model of atopic dermatitis, we demonstrate a pathologic role for CCR8 and mouse CCL8, but not for the TH2-associated chemokine receptor CCR4 or the previously known mouse CCR8 chemokine ligand mouse CCL1 (TCA3), in mediating chronic cutaneous allergic inflammation.

  • 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 Mouse MCP2 (CCL8) Capture Antibody 1 x 600µl
    10X Mouse MCP2 (CCL8) Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CP 1 x 6ml
    Mouse MCP2 (CCL8) 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 factor that attracts monocytes, lymphocytes, basophils and eosinophils. May play a role in neoplasia and inflammatory host responses. This protein can bind heparin. The processed form MCP-2(6-76) does not show monocyte chemotactic activity, but inhibits the chemotactic effect most predominantly of CCL7, and also of CCL2 and CCL5 and CCL8.
  • Tissue specificity

    Highest expression found in the small intestine and peripheral blood cells. Intermediate levels seen in the heart, placenta, lung, skeletal muscle, thymus, colon, ovary, spinal cord and pancreas. Low levels seen in the brain, liver, spleen and prostate.
  • Sequence similarities

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

    N-terminal processed form MCP-2(6-76) is produced by proteolytic cleavage after secretion from peripheral blood monocytes.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Alternative names

    • C C motif chemokine 8
    • CCL 8
    • Ccl8
    • CCL8_HUMAN
    • Chemokine (C C motif) ligand 8
    • Chemokine ligand 8
    • HC14
    • MCP 2
    • MCP-2
    • MCP-2(6-76)
    • Monocyte chemoattractant protein 2
    • Monocyte chemotactic protein 2
    • SCYA10
    • SCYA8
    • Small inducible cytokine A8
    • Small inducible cytokine subfamily A (Cys Cys) member 8
    • Small-inducible cytokine A8
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab203366 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.

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

  • The 1X dilution was 0.025% for the mouse serum and all of the mouse plasmas. Interpolated data values (mean +/- SD, n = 2) are graphed.

  • Titration of mouse stimulated (100 ng/mL LPS + 50 ng/mL PMA, 3 days (+)) lung supernatant, unstimulated (-) lung supernatant, stimulated (5 µg/mL LPS, 6 Days (+)) spleen supernatant, and unstimulated (-) spleen supernatant. A blank cell culture media control was also analyzed, and was negative. The 1X dilution was 10.0% for the stimulated supernatants, and 20.0% for the unstimulated supernatants. Interpolated data values (mean +/- SD, n = 2) are graphed.

Protocols

References

ab203366 has not yet been referenced specifically in any publications.

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