Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-MIF antibody [EPR18149-128] - BSA and Azide free (ab226166)

Overview

  • Product name

    Anti-MIF antibody [EPR18149-128] - BSA and Azide free
    See all MIF primary antibodies
  • Description

    Rabbit monoclonal [EPR18149-128] to MIF - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein within Mouse MIF aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P34884

  • Positive control

    • ICC/IF: Neuro-2a cells.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

    ab226166 is a PBS-only buffer format of ab187064. Please refer to ab187064 for recommended dilutions, protocols, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

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

    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

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18149-128
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab226166 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
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 13 kDa (predicted molecular weight: 13 kDa).
ICC/IF Use at an assay dependent concentration.

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

  • All lanes : Anti-MIF antibody [EPR18149-128] (ab187064) at 1/1000 dilution

    Lane 1 : Wild type HAP1 whole cell lysate
    Lane 2 : MIF knockout HAP1 whole cell lysate
    Lane 3 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
    Lane 4 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate

    Lysates/proteins at 20 µg/ml per lane.

    Predicted band size: 13 kDa



    Ab187064 was shown to specifically react with MIF in wild-type HAP1 cells as signal was lost in MIF knockout cells. Wild-type and MIF knockout samples were subjected to SDS-PAGE. ab187064 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187064).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling MIF with ab187064 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on RAW 264.7 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187064).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Neuro-2a (mouse neuroblastoma cell line) cell line labeling MIF with ab187064 at 1/500 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187064).

  • Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line labeling MIF with ab187064 at 1/500 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187064).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling MIF with ab187064 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Neuro-2a cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab187064).

References

ab226166 has not yet been referenced specifically in any publications.

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