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  1. Link

    products/elisa/human-mcp-1-elisa-kit-ab179886.pdf

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RecombinantSimpleStep

Human MCP-1 ELISA Kit (ab179886)

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  • Protocol Booklet
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Other - Human MCP-1 ELISA Kit (ab179886)
  • Example of human MCP1 standard curve in Sample Diluent NS.
  • Standard curve
  • Interpolated concentrations of native MCP1 in human serum and plasma samples.
  • Interpolated concentrations of native MCP1 in human cerebrospinal fluid, urine, and cell culture supernatant samples.
  • Serum from nine individual healthy human male donors was measured in duplicate.
  • Linearity of dilution.
  • Sandwich ELISA - Human MCP-1 ELISA Kit (ab179886)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 1.26 pg/ml
  • Range: 4.7 pg/ml - 300 pg/ml
  • Sample type: Cell culture supernatant, Cerebral Spinal Fluid, Cit plasma, EDTA Plasma, Hep Plasma, Plasma, Serum, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human MCP-1 ELISA Kit
    See all MCP1 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 2.5%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 6.7%
  • Sample type

    Cell culture supernatant, Urine, Serum, Plasma, Hep Plasma, EDTA Plasma, Cit plasma, Cerebral Spinal Fluid
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    1.26 pg/ml
  • Range

    4.7 pg/ml - 300 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 98 97% - 99%
    Urine 92 90% - 94%
    Serum 97 94% - 101%
    Hep Plasma 91 91% - %
    EDTA Plasma 94 90% - 97%
    Cit plasma 92 91% - 95%
    Cerebral Spinal Fluid 105 102% - 110%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Cow
  • Product overview

    Human MCP1 (CCL2) ELISA kit (ab179886) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of MCP1 protein in human serum, plasma, urine, and cell culture supernatant samples. It uses our proprietary SimpleStep ELISA® technology. Quantitate human MCP1 with 1.26 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 SimpeStep ELISA® kits.


    ASSAY SPECIFICITY This kit recognizes both native and recombinant human MCP1 protein in serum, plasma, urine, and cell culture supernatant samples only. Cell and tissue extract samples have not been tested with this kit.


    SPECIES REACTIVITY This kit recognizes human MCP1 protein.
    Other species reactivity was determined by measuring MCP1 (100%) serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution. 
    Reactivity < 3% was determined for the following species: Mouse and cow.


    CROSS REACTIVITY Recombinant mouse MCP1 and rat MCP1 were prepared at 100 pg/mL and 50 pg/mL and assayed for cross reactivity. Cross-reactivity was only observed at 50 ng/mL.


    CALIBRATION


    This immunoassay is calibrated against a highly purified human MCP1. The NIBSC/WHO unclassified purified human MCP1 preparation 92/794 was evaluated in this kit.


    The dose response curve of the unclassified standard MCP1 parallels the SimpleStep standard curve. To convert sample values obtained with the SimpleStep human MCP1 kit to approximate NIBSC 92/794 units, use the equation below.


    NIBSC (92/794) approximate value (U/mL) = 0.000266 x SimpleStep human MCP1 value (pg/mL).


     

  • Notes

    MCP-1 (CCL2) is a chemotactic factor that attracts monocytes and basophils but not neutrophils or eosinophils. Augments monocyte anti-tumor activity. Has been implicated in the pathogenesis of diseases characterized by monocytic infiltrates, like psoriasis, rheumatoid arthritis or atherosclerosis. May be involved in the recruitment of monocytes into the arterial wall during the disease process of atherosclerosis.

  • 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 Human MCP-1 Capture Antibody 1 x 600µl
    10X Human MCP-1 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Human MCP-1 Lyophilized Recombinant Protein (ab9670) 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
    • Alpha Chemokines (CXC)
    • Immunology
    • Innate Immunity
    • Chemokines
    • Beta Chemokines (CC)
    • Cardiovascular
    • Angiogenesis
    • Angiogenic Factors
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Chemokines
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cytokines and cytokine receptors ELISA kits
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Heart disease
    • Neuroscience
    • Processes
  • Function

    Chemotactic factor that attracts monocytes and basophils but not neutrophils or eosinophils. Augments monocyte anti-tumor activity. Has been implicated in the pathogenesis of diseases characterized by monocytic infiltrates, like psoriasis, rheumatoid arthritis or atherosclerosis. May be involved in the recruitment of monocytes into the arterial wall during the disease process of atherosclerosis.
  • Sequence similarities

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

    Processing at the N-terminus can regulate receptor and target cell selectivity. Deletion of the N-terminal residue converts it from an activator of basophil to an eosinophil chemoattractant.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P13500 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • C-C motif chemokine 2
    • CCL2
    • CCL2_HUMAN
    • Chemokine (C C motif) ligand 2
    • GDCF 2
    • GDCF-2
    • GDCF2
    • HC11
    • HSMCR30
    • JE
    • MCAF
    • MCP 1
    • MCP-1
    • MCP1
    • MGC9434
    • Monocyte chemoattractant protein 1
    • Monocyte chemotactic and activating factor
    • Monocyte chemotactic protein 1
    • Monocyte chemotactic protein-1
    • Monocyte secretory protein JE
    • SCYA2
    • Small inducible cytokine A2
    • Small inducible cytokine A2 (monocyte chemotactic protein 1, homologous to mouse Sig je)
    • Small inducible cytokine subfamily A (Cys Cys), member 2
    • Small-inducible cytokine A2
    • SMC CF
    • SMC-CF
    • SMCCF
    see all
  • Database links

    • Entrez Gene: 6347 Human
    • Omim: 158105 Human
    • SwissProt: P13500 Human
    • Unigene: 303649 Human

    Associated products

    • Alternative Versions

      • Human MCP1 Antibody Pair - BSA and Azide free (ab241803)

    Images

    • Other - Human MCP-1 ELISA Kit (ab179886)
      Other - Human MCP-1 ELISA Kit (ab179886)

      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 human MCP1 standard curve in Sample Diluent NS.
      Example of human MCP1 standard curve in Sample Diluent NS.

      The MCP1 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Standard curve
      Standard curve

      Example of human MCP1 standard curve in Sample Diluent NS. The MCP1 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of native MCP1 in human serum and plasma samples.
      Interpolated concentrations of native MCP1 in human serum and plasma samples.

      The concentrations of MCP1 were measured in duplicates, interpolated from the MCP1 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 100%, plasma (citrate) 25%, plasma (heparin), 25%, and plasma (EDTA). The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MCP1 concentration was determined to be 72 pg/mL in serum, 96 pg/mL in plasma (citrate), 101 pg/mL in plasma (heparin), and 88 pg/mL plasma (EDTA).

    • Interpolated concentrations of native MCP1 in human cerebrospinal fluid, urine, and cell culture supernatant samples.
      Interpolated concentrations of native MCP1 in human cerebrospinal fluid, urine, and cell culture supernatant samples.

      The concentrations of MCP1 were measured in duplicates, interpolated from the MCP1 standard curves and corrected for sample dilution. Undiluted samples are as follows: cerebrospinal fluid (50%)), urine 100%, and PHA stimulated PBMC supernatant 0.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MCP1 concentration was determined to be 371 pg/mL in cerebrospinal fluid, 144 pg/mL in urine, and 70217 pg/mL in PHA stimulated PBMC supernatant.

    • Serum from nine individual healthy human male donors was measured in duplicate.
      Serum from nine individual healthy human male donors was measured in duplicate.

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MCP1 concentration was determined to be 113 pg/mL with a range of 58 - 247 pg/mL.

    • Linearity of dilution.
      Linearity of dilution.

      Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.

      Native MCP1 was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.

    • Sandwich ELISA - Human MCP-1 ELISA Kit (ab179886)
      Sandwich ELISA - Human MCP-1 ELISA Kit (ab179886)
      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 (17)

    Publishing research using ab179886? Please let us know so that we can cite the reference in this datasheet.

    ab179886 has been referenced in 17 publications.

    • Pedrosa MA  et al. Experimental data using candesartan and captopril indicate no double-edged sword effect in COVID-19. Clin Sci (Lond) 135:465-481 (2021). PubMed: 33479758
    • Bengali M  et al. Non-invasive intradermal imaging of cystine crystals in cystinosis. PLoS One 16:e0247846 (2021). PubMed: 33661986
    • Das NA  et al. Empagliflozin reduces high glucose-induced oxidative stress and miR-21-dependent TRAF3IP2 induction and RECK suppression, and inhibits human renal proximal tubular epithelial cell migration and epithelial-to-mesenchymal transition. Cell Signal 68:109506 (2020). PubMed: 31862399
    • Hosseinkhani B  et al. (Sub)populations of extracellular vesicles released by TNF-a -triggered human endothelial cells promote vascular inflammation and monocyte migration. J Extracell Vesicles 9:1801153 (2020). PubMed: 32944190
    • Zhang H  et al. Long noncoding RNA MIAT2 alleviates lipopolysaccharide-induced inflammatory damage in WI-38 cells by sponging microRNA-15. J Cell Physiol 235:3690-3697 (2020). PubMed: 31566734
    View all Publications for this product

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