The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 19066630
Use at an assay dependent concentration.
Use at an assay dependent concentration.
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.
Belongs to the MIF family.
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.
Anti-MIF antibody (ab7207) + LPS stimulated RAW cell lysate
Performed under reducing conditions.
MIF detected by Western
Blot in LPS stimulated
RAW cell lysate.
Immunohistochemistry (Frozen sections) - Anti-MIF antibody (ab7207)This image is courtesy of an anonymous Abreview
ab7207 staining MIF in Mouse small intestine tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with Dako protein block for 1 hour at room temperature. Samples were incubated with primary antibody (1/200) for 1 hour. An undiluted Alkaline Phosphatase-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIF antibody (ab7207)This image is courtesy of an anonymous Abreview
ab7207 staining MIF in Rat liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% protein blocking solution for 1 hour at room temperature; antigen retrieval was by heat mediation with a citrate EDTA buffer (pH 6.2). Samples were incubated with primary antibody (1/200 in antibody diluent) for 1 hour. An undiluted Alkaline phosphatase-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-MIF antibody (ab7207)This image is courtesy of an Abreview submitted by Dr James Harris
ab7207 staining MIF in murine immortalised bone marrow-derived macrophages by Immunocytochemistry/ Immunofluorescence.
Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 5% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/250 in 5% goat serum/3% BSA in PBS) for 1 hour at 20°C. An AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG (1/500) was used as the secondary antibody.
Western blot - Anti-MIF antibody (ab7207)Image courtesy of an anonymous Abreview.
All lanes : Anti-MIF antibody (ab7207) at 1/1000 dilution
Lanes 1-2 : Whole cell lysate prepared from normal rat serum Lanes 3-4 : Whole cell lysate prepared from rat serum (90 minutes warm ischemia and 1h reperfusion)
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Immunohistochemistry (Frozen sections) - Anti-MIF antibody (ab7207)This image is courtesy of an Abreview submitted by Miss Silke Vorwald
ab7207 staining mouse Testis tissue sections by Immunohistochemistry (Frozen sections). Tissue was fixed in paraformaldehyde and blocked in 1% serum for 10 minutes at 20°C. The primary antibody was diluted 1/100 and incubated with sample for 20 hours at 20°C. A biotin labelled goat polyclonal to rabbit IgG antibody, diluted 1/500 was used as the secondary.
ELISA - Anti-MIF antibody (ab7207)Image courtesy of an anonymous Abreview.
ab7207 used at a 1/1000 dilution as detection antibody in ELISA testing MIF in rat serum samples. Samples were blocked with 1% BSA for 2 hours at room temperature. The secondary used was an HRP conjugated donkey anti-goat (H+L) polyclonal used at a 1/200,000 dilution.
Korf H et al. MIF inhibition interferes with the inflammatory and T cell-stimulatory capacity of NOD macrophages and delays autoimmune diabetes onset. PLoS One12:e0187455 (2017).
Read more (PubMed: 29095944) »
Fex Svenningsen Å et al. Macrophage migration inhibitory factor (MIF) modulates trophic signaling through interaction with serine protease HTRA1. Cell Mol Life Sci74:4561-4572 (2017).
Read more (PubMed: 28726057) »