Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-MIF antibody [EPR12463] - BSA and Azide free (ab183302)

Overview

  • Product name

    Anti-MIF antibody [EPR12463] - BSA and Azide free
    See all MIF primary antibodies
  • Description

    Rabbit monoclonal [EPR12463] to MIF - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • WB: HAP1, HeLa and HepG2 cell lysates.
  • General notes

    Ab183302 is the carrier-free version of ab175189. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab183302 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

    EPR12463
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab183302 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 Use at an assay dependent concentration. Predicted molecular weight: 12 kDa.

 

IP Use at an assay dependent concentration.

 

Flow Cyt Use at an assay dependent concentration.

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

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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

  • 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab175189).

References

ab183302 has not yet been referenced specifically in any publications.

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