Human MIF Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human MIF in direct ELISAs.
  • Source
    Monoclonal Mouse IgG1 Clone # 932606
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human MIF
    Met1-Phe114
    Accession # P14174
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    THP‑1 human acute monocytic leukemia cell line and U937 human histiocytic lymphoma cell line with 2 µg/mL of Mouse Anti-Human MIF Monoclonal Antibody (Catalog # MAB2891). It is recommended to use Mouse Anti-Human MIF Monoclonal Antibody (Catalog # MAB2893, Clone # 932603) as an alternative antibody for Western blot.
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human MIF by Western Blot. Western blot shows lysates of THP‑1 human acute monocytic leukemia cell line and U937 human histiocytic lymphoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human MIF Monoclonal Antibody (Catalog # MAB2891) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for MIF at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1. It is recommended to use Mouse Anti-Human MIF Monoclonal Antibody (Catalog # MAB2893, Clone # 932603) as an alternative antibody for Western blot.
Intracellular Staining by Flow Cytometry
Detection of MIF in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cell (PBMCs), resting (open histogram), or treated with 1 ug/mL LPS overnight and 3 uM monensin for 2 hours (filed histogram) were stained with Mouse Anti-Human MIF Monoclonal Antibody (Catalog # MAB2891) or isotype control antibody (Catalog # MAB002), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: MIF

MIF (or macrophage Migration Inhibitory Factor) was the first lymphokine/cytokine to be recognized in the pregenomics era (1, 2). Regardless, it is one of the least understood of all inflammatory mediators (1, 3). Human MIF is a 12.5 kDa, 115 amino acid (aa) nonglycosylated polypeptide that is synthesized without a signal sequence (4-7). Secretion occurs nonclassically via an ABCA1 transporter (8). The initiating Met is removed, leaving Pro as the first amino acid. The molecule consists of two alpha -helices and six beta -strands, four of which form a beta -sheet. The two remaining beta -strands interact with other MIF molecules, creating a trimer (2, 9, 10). Structure-function studies suggest MIF is bifunctional with segregated topology. The N- and C-termini mediate enzyme activity (in theory). Phenylpyruvate tautomerase activity (enol-to-keto) has been demonstrated and is dependent upon Pro at position #1 (11). Amino acids 50-65 have also been suggested to contain thiol-protein oxidoreductase activity (12). MIF has proinflammatory cytokine activity centered around aa’s 49-65. On fibroblasts, MIF induces, IL-1, IL-8, and MMP expression; on macrophages, MIF stimulates NO production and TNF-alpha release following IFN-gamma activation (13, 14). MIF apparently acts through CD74 and CD44, likely in some form of trimeric interaction (15, 16). Human MIF is active on mouse cells (14). Human MIF is 90%, 94%, 95%, and 90% aa identical to mouse, bovine, porcine, and rat MIF, respectively.

  • References:
    1. Norand, E.F. and M. Leech (2005) Front. Biosci. 10:12.
    2. Donn, R.P. and D.W. Ray (2004) J. Endocrinol. 182:1.
    3. Calandra, T. and T. Roger (2003) Nat. Rev. Immunol. 3:791.
    4. Kozak, C.A. et al. (1995) Genomics 27:405.
    5. Weiser, W.Y. et al. (1989) Proc. Natl. Acad. Sci. USA 86:7522.
    6. Paralkar, V. and G. Wistow (1994) Genomics 19:48.
    7. Wistow, G.J. et al. (1993) Proc. Natl. Acad. Sci. USA 90:1272.
    8. Flieger, O. et al. (2003) FEBS Lett. 551:78.
    9. Philo, J.S. et al. (2004) Biophys. Chem. 108:77.
    10. Sun, H-W. et al. (1996) Protein Eng. 9:631.
    11. Stamps, S.L. et. al. (2000) Biochemistry 39:9671.
    12. Nguyen, M.T. et al. (2003) J. Biol. Chem. 278:33654.
    13. Sato, A. et al. (2003) Dev. Comp. Immunol. 27:401.
    14. Bernhagen, J. et al. (1994) Biochemistry 33:14144.
    15. Leng, L. et al. (2003) J. Exp. Med. 197:1467.
    16. Meyer-Siegler, K.L. and P.L. Vera (2005) J. Urol. 173:615.
  • Long Name:
    Macrophage Migration Inhibitory Factor
  • Entrez Gene IDs:
    4282 (Human); 17319 (Mouse)
  • Alternate Names:
    EC 5.3.2.1; EC 5.3.3.12; GIFmacrophage migration inhibitory factor; GLIF; Glycosylation-inhibiting factor; L-dopachrome isomerase; L-dopachrome tautomerase; macrophage migration inhibitory factor (glycosylation-inhibiting factor); MIF; MMIF; Phenylpyruvate tautomerase
Related Research Areas
Isotype Controls
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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse IgG Antibody

WB AF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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