Overview

  • Product name
    Anti-MIF antibody [EPR12463]
    See all MIF primary antibodies
  • Description
    Rabbit monoclonal [EPR12463] to MIF
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, Flow Cytmore details
    Unsuitable for: ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human MIF aa 50 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
    Database link: P14174

  • Positive control
    • WB: Y79, THP-1 and Molt-4 cell lysates. Mouse and rat brain tissue lysates. Flow Cyt: THP-1 and Molt-4 cells. IP: Human fetal brain tissue lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR12463
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab175189 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
WB 1/1000 - 1/10000. Predicted molecular weight: 12 kDa.

For unpurified use at 1/500 - 1/1000.

IP 1/50.

For unpurified use at 1/30.

Flow Cyt 1/140.

For unpurified use at 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • 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.
    • Information by UniProt
    • Database links
    • Alternative names
      • GIF antibody
      • GLIF antibody
      • Glycosylation inhibiting factor antibody
      • Glycosylation-inhibiting factor antibody
      • L-dopachrome isomerase antibody
      • L-dopachrome tautomerase antibody
      • Macrophage migration inhibitory factor (glycosylation-inhibiting factor) antibody
      • Macrophage migration inhibitory factor antibody
      • MIF antibody
      • MIF protein antibody
      • MIF_HUMAN antibody
      • MMIF antibody
      • Phenylpyruvate tautomerase antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: MIF knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: HepG2 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab175189 observed at 12 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab175189 was shown to specifically react with MIF in wild-type HAP1 cells along with additional cross-reactive bands. No bands were observed when knockout cells were examined. Wild-type and MIF knockout samples were subjected to SDS-PAGE. Ab175189 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/2,0000 dilution for 1 hour at room temperature before imaging.

    • Flow cytometry analysis of THP-1 cells labelling MIF with unpurified ab175189 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary anitbody. A rabbit monoclonal IgG was used as the isotype control (green).

    • All lanes : Anti-MIF antibody [EPR12463] (ab175189) at 1/1000 dilution (unpurified)

      Lane 1 : Y79 cell lysate
      Lane 2 : Mouse brain tissue lysate
      Lane 3 : Rat brain tissue lysate

      Lysates/proteins at 20 µg/ml per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 12 kDa
      Observed band size: 12 kDa



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-MIF antibody [EPR12463] (ab175189) at 1/2000 dilution (purified)

      Lane 1 : Y79 cell lysate
      Lane 2 : Mouse brain tissue lysate
      Lane 3 : Rat brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 12 kDa
      Observed band size: 12 kDa



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-MIF antibody [EPR12463] (ab175189) at 1/1000 dilution (unpurified)

      Lane 1 : Y79 lysates
      Lane 2 : THP-1 lysates
      Lane 3 : Molt-4 lysates
      Lane 4 : Human fetal brain lysates

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 12 kDa

    • Flow cytometry analysis of THP-1 cells labelling MIF with purified ab175189 at 1/140 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary anitbody. A rabbit monoclonal IgG was used as the isotype control (green).

    • Flow cytometric analysis of permeabilized Molt-4 cells using unpurified ab175189 at a 1/10 dilution (red) or a rabbit IgG (negative) (green).

    • ab175189 (unpurified) at 1/30 immunoprecipitating MIF in human fetal brain. For western blotting, a Peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • ab175189 (purified) at 1/50 immunoprecipitating MIF in human fetal brain. For western blotting, a Peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    References

    This product has been referenced in:
    • Kim JH  et al. NADPH oxidase 4 is required for the generation of macrophage migration inhibitory factor and host defense against Toxoplasma gondii infection. Sci Rep 7:6361 (2017). Read more (PubMed: 28743960) »
    • Jin Y & Dai Z USO1 promotes tumor progression via activating Erk pathway in multiple myeloma cells. Biomed Pharmacother 78:264-71 (2016). WB ; Human . Read more (PubMed: 26898451) »

    See all 4 Publications for this product

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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