Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-MIF antibody [EPR12462] (ab176565)

Overview

  • Product name

    Anti-MIF antibody [EPR12462]
    See all MIF primary antibodies
  • Description

    Rabbit monoclonal [EPR12462] to MIF
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Sheep, Chicken, Cow, Cynomolgus monkey, Rhesus monkey
  • Immunogen

    Synthetic peptide within Human MIF aa 1-100 (Cysteine residue). The exact sequence is proprietary.
    Database link: P14174

  • Positive control

    • Y79, Jurkat, A375 and THP1 cell lysates.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR12462
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab176565 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.
IP 1/10 - 1/100.

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 - ab176565 observed at 12 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab176565 was shown to specifically recognize MIF in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when knockout cells were examined. Wild-type and MIF knockout samples were subjected to SDS-PAGE. Ab176565 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 1/20,000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-MIF antibody [EPR12462] (ab176565) at 1/1000 dilution

    Lane 1 : Y79 cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : A375 cell lysate
    Lane 4 : THP1 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP at 1/2000 dilution

    Developed using the ECL technique.

    Predicted band size: 12 kDa

  • Western blot analysis on immunoprecipitation pellet from Jurkat cell lysate labeling MIF with ab176565 at 1/10 dilution.

References

ab176565 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.1% saponin
Specification
HeLa
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 37°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 09 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing (13)
Loading amount
20 cells
Specification
HeLa
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 14 2019

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