Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

Western Blot, Intracellular Staining by Flow Cytometry, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 932606
View all MIF Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human MIF
Met1-Phe114
Accession # P14174

Specificity

Detects human MIF in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human MIF Antibody

Detection of Human MIF antibody by Western Blot.

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.
Detection of MIF antibody in Human PBMCs antibody 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).

Applications for Human MIF Antibody

Application
Recommended Usage

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
Sample: Human peripheral blood mononuclear cell (PBMCs) treated with LPS overnight and monensin for 2 hours were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).

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.

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

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), MMIF, Phenylpyruvate tautomerase

Entrez Gene IDs

4282 (Human); 17319 (Mouse)

Gene Symbol

MIF

UniProt

P14174

Additional MIF Products

Product Documents for Human MIF Antibody

Certificate of Analysis

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Product Specific Notices for Human MIF Antibody

For research use only

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