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
Product nameHRP Anti-MIF antibody [EPR12463]
See all MIF primary antibodies
DescriptionHRP Rabbit monoclonal [EPR12463] to MIF
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
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
- WB: Y70 whole cell lysate and Human Fetal Brain tissue lysate. IHC-P: Normal human lung tissue.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: 30% Glycerol, 1% BSA, PBS
Concentration information loading...
PurityProtein A purified
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.
|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).|
FunctionPro-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 diseaseGenetic 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 similaritiesBelongs to the MIF family.
Cellular localizationSecreted. 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
- GIF antibody
- GLIF antibody
- Glycosylation inhibiting factor antibody
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.
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.
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.
ab196645 has not yet been referenced specifically in any publications.