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    mouse-mif-elisa-kit-ab209885.pdf

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Immunology Innate Immunity Macrophage / Inflamm.
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RecombinantSimpleStep

Mouse MIF ELISA Kit (ab209885)

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Other - Mouse MIF ELISA Kit (ab209885)
  • Example of mouse MIF standard curve in Sample Diluent NS
  • Example of mouse MIF standard curve in 1X Extraction Buffer PTR
  • Interpolated concentrations of native MIF in mouse serum, plasma and cell culture supernatant samples
  • Interpolated concentrations of native MIF in RAW 264.7 cell extract and mouse spleen based on 0.49 µg/mL and 3.91 µg /mL extract loads, respectively
  • Sandwich ELISA - Mouse MIF ELISA Kit (ab209885)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 28 pg/ml
  • Range: 125 pg/ml - 8000 pg/ml
  • Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Tissue Extracts, Tissue Homogenate
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Mouse

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Overview

  • Product name

    Mouse MIF ELISA Kit
    See all MIF kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Mouse serum 5 13.6%
    Inter-assay
    Sample n Mean SD CV%
    Mouse serum 3 9.8%
  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma, Tissue Homogenate
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    28 pg/ml
  • Range

    125 pg/ml - 8000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 110 106% - 117%
    Cell culture extracts 111 108% - 113%
    Tissue Extracts 108 102% - 118%
    Hep Plasma 112 105% - 117%
    EDTA Plasma 109 105% - 112%
    Cit plasma 112 112% - 113%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Mouse MIF ELISA Kit (ab209885) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of MIF protein in cell culture extracts, cell culture supernatant, cit plasma, edta plasma, hep plasma, serum, tissue extracts, and tissue homogenate. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse MIF with 28 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

    Macrophage migration inhibitory factor (MIF) is a 12.37 kDa pro-inflammatory cytokine and is involved in the innate immune response to bacterial pathogens. MIF assembles into a trimer, composed of three identical subunits. The expression of MIF at sites of inflammation suggests a role as mediator in regulating the function of macrophages in host defense. MIF counteracts the anti-inflammatory activity of glucocorticoids and exhibits phenylpyruvate tautomerase and dopachrome tautomerase activity. It is not clear whether the tautomerase activity has any physiological relevance, and whether it is important for cytokine activity.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • 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 MIF Capture Antibody 1 x 600µl
    10X Mouse MIF 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 4BR 1 x 6ml
    Mouse MIF 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
    • Macrophage / Inflamm.
  • Function

    Pro-inflammatory cytokine. Involved in the innate immune response to bacterial pathogens. The expression of MIF at sites of inflammation suggests a role as mediator in regulating the function of macrophages in host defense. Counteracts the anti-inflammatory activity of glucocorticoids. Has phenylpyruvate tautomerase and dopachrome tautomerase activity (in vitro), but the physiological substrate is not known. It is not clear whether the tautomerase activity has any physiological relevance, and whether it is important for cytokine activity.
  • Involvement in disease

    Genetic variations in MIF are associated with susceptibility to rheumatoid arthritis systemic juvenile (RASJ) [MIM:604302]. An inflammatory articular disorder with systemic-onset beginning before the age of 16. It represents a subgroup of juvenile arthritis associated with severe extraarticular features and occasionally fatal complications. During active phases of the disorder, patients display a typical daily spiking fever, an evanescent macular rash, lymphadenopathy, hepatosplenomegaly, serositis, myalgia and arthritis.
  • Sequence similarities

    Belongs to the MIF family.
  • Cellular localization

    Secreted. Cytoplasm. Does not have a cleavable signal sequence and is secreted via a specialized, non-classical pathway. Secreted by macrophages upon stimulation by bacterial lipopolysaccharide (LPS), or by M.tuberculosis antigens.
  • Target information above from: UniProt accession P14174 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • GIF
    • GLIF
    • Glycosylation inhibiting factor
    • Glycosylation-inhibiting factor
    • L-dopachrome isomerase
    • L-dopachrome tautomerase
    • Macrophage migration inhibitory factor
    • Macrophage migration inhibitory factor (glycosylation-inhibiting factor)
    • MIF
    • MIF protein
    • MIF_HUMAN
    • MMIF
    • Phenylpyruvate tautomerase
    see all
  • Database links

    • Entrez Gene: 17319 Mouse
    • SwissProt: P34884 Mouse
    • Unigene: 2326 Mouse

    Associated products

    • ELISA kits

      • Mouse MIF Matched Antibody Pair Kit (ab215080)

    Images

    • Other - Mouse MIF ELISA Kit (ab209885)
      Other - Mouse MIF ELISA Kit (ab209885)

      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 mouse MIF standard curve in Sample Diluent NS
      Example of mouse MIF standard curve in Sample Diluent NS

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

    • Example of mouse MIF standard curve in 1X Extraction Buffer PTR
      Example of mouse MIF standard curve in 1X Extraction Buffer PTR

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

    • Interpolated concentrations of native MIF in mouse serum, plasma and cell culture supernatant samples
      Interpolated concentrations of native MIF in mouse serum, plasma and cell culture supernatant samples

      The concentrations of MIF were measured in duplicates, interpolated from the MIF standard curves and corrected for sample dilution. Undiluted samples are as follows:  serum 0.5%, plasma (citrate) 2%, plasma (EDTA) 2%, plasma (heparin) 2%, and stimulated RAW 264.7 cell culture supernatant 0.3%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean MIF concentration was determined to be 1.06 µg/mL in serum, 286 ng/mL in plasma (citrate), 229 ng/mL in plasma (EDTA), 293 ng/mL in plasma (heparin), and 1.75 µg/mL in LPS-stimulated RAW 264.7 cell culture supernatant.

    • Interpolated concentrations of native MIF in RAW 264.7 cell extract and mouse spleen based on 0.49 µg/mL and 3.91 µg /mL extract loads, respectively
      Interpolated concentrations of native MIF in RAW 264.7 cell extract and mouse spleen based on 0.49 µg/mL and 3.91 µg /mL extract loads, respectively

      The concentrations of MIF were measured in duplicate and interpolated from the MIF standard curve and corrected for sample dilution.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).  The mean MIF concentration was determined to be 9.8 µg/mL in RAW 264.7 cell extract and 1.1 µg/mL in mouse spleen extract.

    • Sandwich ELISA - Mouse MIF ELISA Kit (ab209885)
      Sandwich ELISA - Mouse MIF ELISA Kit (ab209885)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (0)

    Publishing research using ab209885? Please let us know so that we can cite the reference in this datasheet.

    ab209885 has not yet been referenced specifically in any publications.

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