MIF Antibody (932606) [Unconjugated]

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.

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).

• Reactivity: Hu
• Applications: WB, CyTOF-ready, Flow
• Clone: 932606
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

MIF Antibody (932606) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human MIF
  Met1-Phe114
  Accession # P14174
• Specificity: Detects human MIF in direct ELISAs.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: MIF
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for MIF Antibody (932606) [Unconjugated]

• 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

Background

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.

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: MIF
• Uniprot:
  • P14174()