Validated using a knockout cell lineRecombinantRabMAb

Recombinant HRP Anti-MIF antibody [EPR12463] (ab196645)

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Western blot - HRP Anti-MIF antibody [EPR12463] (ab196645)
  • Western blot - HRP Anti-MIF antibody [EPR12463] (ab196645)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-MIF antibody [EPR12463] (ab196645)
  • HRP Anti-MIF antibody [EPR12463] (ab196645)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • HRP Rabbit monoclonal [EPR12463] to MIF
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human
  • Conjugation: HRP

Overview

  • Product name

    HRP Anti-MIF antibody [EPR12463]
    See all MIF primary antibodies

  • Description

    HRP Rabbit monoclonal [EPR12463] to MIF

  • Host species

    Rabbit

  • Conjugation

    HRP

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • 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: Y70 whole cell lysate and Human Fetal Brain tissue lysate. IHC-P: Normal human lung tissue.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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. Avoid freeze / thaw cycle. Store In the Dark.

  • Storage buffer

    pH: 7.40
    Preservative: 0.1% 10% Proclin 300 Solution
    Constituents: 30% Glycerol, 1% BSA, PBS
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR12463

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab196645 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
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 12 kDa (predicted molecular weight: 12 kDa).

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

    see all

  • 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

  • Western blot - HRP Anti-MIF antibody [EPR12463] (ab196645)
    Western blot - HRP Anti-MIF antibody [EPR12463] (ab196645)
    All lanes : HRP Anti-MIF antibody [EPR12463] (ab196645) at 2 µg/ml

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : MIF knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

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



    ab196645 was shown to recognize MIF (Macrophage migration inhibitory factor) in wild-type HAP1 cells as signal was lost at the expected MW in MIF knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and MIF knockout samples were subjected to SDS-PAGE. Ab196645 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 2 μg/ml and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • Western blot - HRP Anti-MIF antibody [EPR12463] (ab196645)
    Western blot - HRP Anti-MIF antibody [EPR12463] (ab196645)
    All lanes : HRP Anti-MIF antibody [EPR12463] (ab196645) at 1/2000 dilution

    Lane 1 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
    Lane 2 : Brain (Human) Tissue Lysate - fetal normal tissue (ab29467)

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 4 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab196645 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-MIF antibody [EPR12463] (ab196645)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-MIF antibody [EPR12463] (ab196645)

    IHC image of MIF staining in a section of formalin-fixed paraffin-embedded normal human lung, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab196645 at 1/500 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • HRP Anti-MIF antibody [EPR12463] (ab196645)
    HRP Anti-MIF antibody [EPR12463] (ab196645)

References (0)

Publishing research using ab196645? Please let us know so that we can cite the reference in this datasheet.

ab196645 has not yet been referenced specifically in any publications.

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