Human MIF Antibody

Catalog #: MAB2891 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB2891
MAB2891-SP

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Human MIF Antibody Summary

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 either lyophilized or 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.

Scientific Data

Western Blot

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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 antibody in Human PBMCs antibody by Flow Cytometry.

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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).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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.

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

Norand, E.F. and M. Leech (2005) Front. Biosci. 10:12.
Donn, R.P. and D.W. Ray (2004) J. Endocrinol. 182:1.
Calandra, T. and T. Roger (2003) Nat. Rev. Immunol. 3:791.
Kozak, C.A. et al. (1995) Genomics 27:405.
Weiser, W.Y. et al. (1989) Proc. Natl. Acad. Sci. USA 86:7522.
Paralkar, V. and G. Wistow (1994) Genomics 19:48.
Wistow, G.J. et al. (1993) Proc. Natl. Acad. Sci. USA 90:1272.
Flieger, O. et al. (2003) FEBS Lett. 551:78.
Philo, J.S. et al. (2004) Biophys. Chem. 108:77.
Sun, H-W. et al. (1996) Protein Eng. 9:631.
Stamps, S.L. et. al. (2000) Biochemistry 39:9671.
Nguyen, M.T. et al. (2003) J. Biol. Chem. 278:33654.
Sato, A. et al. (2003) Dev. Comp. Immunol. 27:401.
Bernhagen, J. et al. (1994) Biochemistry 33:14144.
Leng, L. et al. (2003) J. Exp. Med. 197:1467.
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