Macrophage Migration Inhibitory Factor (Glycosylation-Inhibiting Factor) (MIF) antibody

Antigen: Macrophage Migration Inhibitory Factor (Glycosylation-Inhibiting Factor) (MIF)

Synonyms: GIF, GLIF, MMIF, Glif, MGC107654, MGC72801, MIF, MGC127044, mif, Mif, gif, glif, mmif, LOC100136498, LOC100284350, LOC100284546

Clonality: Polyclonal

Host: Chicken

Reactivity: Human

Conjugate: Un-conjugated

Application: ELISA, Western Blotting (WB)

Catalog no.: ABIN118028

Additional Information

Alternative name: MIF

UniProt: P14174

Immunogen: This IgY fraction antibody was prepared from eggs of chickens laid after repeatedimmunizations with a synthetic peptide corresponding to aa 2-32 of Human MIF conjugatedto keyhole limpet hemocyanin (KLH). MIF is a proinflammatory cytokine that plays animportant role in systemic inflammatory events. AA Sequence: P-M-F-I-V-N-T-N-V-P-R-A-S-V-P-D-G-F-L-S-E-L-T-Q-Q-L-A-Q-A-T-G

Cross-Reactivity: Human

Description: Cytokines play an important role in inflammation and immunity. Macrophage migrationinhibitory factor (MIF) was one of the first cytokine activities described and is aproinflammatory cytokine that plays an important role in systemic inflammatory events. MIF's cytokine activity was initially described as a T cell-derived factor that inhibited therandom migration of macrophages, hence its name. Since it was cloned and expressed inpure form, MIF's activities have been established to play many roles in development andvarious disease states. For example, MIF is released from macrophages and T cells inresponse to physiological concentrations of glucocorticoids. The secreted MIFcounterregulates the immunosuppressive effects of steroids on immune cell activation andcytokine production. In in vitro experiments MIF is significantly upregulated by thestimulation of lipopolysaccharide (LPS) using 10 pg/ml to 10 ng/ml of LPS and reaches themaximum 12 h after the stimulation. MIF also prevents cleavage of Bax into an 18-kDaactive fragment, and, consequently, reduces activation of the critical effector caspase 3,suggesting that MIF inhibits apoptosis pathways proximal to mitochondria activation andis therefore a survival factor. And, MIF is also known to exhibit enzymatic activities. Apathologic role for MIF has been described in many conditions including arthritis, asthma,and inflammatory bowel disease, and may also play a role in the control of cell growth incertain cancers. Consequently MIF is suggested to be a potential therapeutic target forhuman diseases.

Characteristics: Synonyms: Macrophage migration inhibitory factor, Phenylpyruvate tautomerase,Glycosylation-inhibiting factor, GLIF, MMIF

Application Details

Application Notes: This IgY fraction antibody of anti-Human MacrophageMigration Inhibitory Factor (MIF) has been tested for use in ELISA and Immunoblotting. Although not tested, this antibody may also be useful for Neutralization assays,Immunohistochemistry and Flow Cytometry. The antibody recognizes 12,300 MW maturehuman MIF. The MIF gene encodesa protein of 115 aa. The initiating methionine is cleaved leaving a mature protein of 114 aa. Reactivity in other immunoassays is unknown. Immunoblot: Using IRDYE800 conjugated Goat-a-Chicken IgG [H&L] MX10. A workingdilution range of 1: 500-1: 1,000 is suggested for this application to detect human MIF fromsupernatants or lysates of 2 x 10e6 endotoxin-stimulated human peripheral bloodmononuclear cells (PBMC). PBMC are stimulated for 24 hours with 1% (v/v) human serumplus 10 ng/mL E. coli LPS. This product has been assayed by ELISA against human MIF using HRP ConjugatedGoat-a-Chicken IgG [H&L] MX10 and ABTS as a substrate for 30 minutes at roomtemperature. A working dilution range of 1: 1,000-1: 2,000 is suggested for this product. For use in ELISAformats, this antibody is best used as the second antibody in combination with amonoclonal antibody as a capture antibody. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Concentration: 5.0 mg/ml (by UV absorbance at 280 nm)

Purification: Purified

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 with 0.01%(w/v) Sodium Azide as preservative.Reconstitution: Restore with 0.1 ml of deionized water (or equivalent).

Storage: Store vial at 2-8°C prior to restoration. Restore with 0. 1 ml of deionized water (orequivalent). For extended storage aliquot contents and freeze at -20°C or below. Centrifugeproduct if not completely clear after standing at room temperature. This product is stablefor one month at 2-8°C as an undiluted liquid. Dilute only prior to immediate use. Avoid cycles of freezing and thawing. Shelf life: one year from despatch.

Research Area: Immunology, Innate Immunity, Cytokines, Inflammation

Restrictions: For Research Use only

Images

Western blot analysis of.,.IgY fraction of Chicken-anti-Human MIF.,.polyclonal antibody shows the.,.detection of 100 ng of recombinant.,.MIF present in a lysate. Similar.,.detection of MIF will occur when.,.human serum is analyzed. In lane 1.,.no reaction is observed in the.,.control whereas lane 2 shows a.,.single band at 12.3 kDa. A 4-20%.,.gradient gel was used to separate.,.the proteins by SDS-PAGE. The.,.protein was transferred to nitrocellulose.,.using standard methods..,.After blocking the membrane was.,.probed with the primary antibody for.,.1 h at room temperature followed by.,.washes and reaction with a 1:5,000.,.dilution of IRDye™800 conjugated.,.Gt-a-Chicken Rabbit IgG [H&L].,.for 1 h at room temperature..,.LICOR's Odyssey® Infrared.,.Imaging System was used.,.to scan and process the image..,.Other detection systems will yield.,.similar results.

Publications

Weiser, Temple, Witek-Giannotti et al.: "Molecular cloning of a cDNA encoding a human macrophage migration inhibitory factor." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 86, Issue 19, pp. 7522-6, 1989 (PubMed).

Calandra, Bernhagen, Metz et al.: "MIF as a glucocorticoid-induced modulator of cytokine production." in: Nature, Vol. 377, Issue 6544, pp. 68-71, 1995 (PubMed).

Mitchell, Bacher, Bernhagen et al.: "Cloning and characterization of the gene for mouse macrophage migration inhibitory factor (MIF)." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 154, Issue 8, pp. 3863-70, 1995 (PubMed).

Bernhagen, Calandra, Mitchell et al.: "MIF is a pituitary-derived cytokine that potentiates lethal endotoxaemia." in: Nature, Vol. 365, Issue 6448, pp. 756-9, 1993 (PubMed).

Claret, Hibi, Dhut et al.: "A new group of conserved coactivators that increase the specificity of AP-1 transcription factors." in: Nature, Vol. 383, Issue 6599, pp. 453-7, 1996 (PubMed).

Rosengren, Bucala, Aman et al.: "The immunoregulatory mediator macrophage migration inhibitory factor (MIF) catalyzes a tautomerization reaction." in: Molecular medicine (Cambridge, Mass.), Vol. 2, Issue 1, pp. 143-9, 1997 (PubMed).

Bernhagen, Calandra, Bucala: "Regulation of the immune response by macrophage migration inhibitory factor: biological and structural features." in: Journal of molecular medicine (Berlin, Germany), Vol. 76, Issue 3-4, pp. 151-61, 1998 (PubMed).

Bucala: "Neuroimmunomodulation by macrophage migration inhibitory factor (MIF)." in: Annals of the New York Academy of Sciences, Vol. 840, pp. 74-82, 1998 (PubMed).

Kleemann, Kapurniotu, Frank et al.: "Disulfide analysis reveals a role for macrophage migration inhibitory factor (MIF) as thiol-protein oxidoreductase." in: Journal of molecular biology, Vol. 280, Issue 1, pp. 85-102, 1998 (PubMed).

Onodera, Kaneda, Mizue et al.: "Macrophage migration inhibitory factor up-regulates expression of matrix metalloproteinases in synovial fibroblasts of rheumatoid arthritis." in: The Journal of biological chemistry, Vol. 275, Issue 1, pp. 444-50, 2000 (PubMed).

Kleemann, Hausser, Geiger et al.: "Intracellular action of the cytokine MIF to modulate AP-1 activity and the cell cycle through Jab1." in: Nature, Vol. 408, Issue 6809, pp. 211-6, 2000 (PubMed).