MIF Antibody [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 0.5 µg/mL of Goat Anti-Human/Mouse/Rat MIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-289-PB) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). 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.

Western blot shows lysates of J774A.1 mouse reticulum cell sarcoma macrophage cell line and NR8383 rat alveolar macrophage cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat MIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-289-PB) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for MIF at approximately 12-14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of THP‑1 human acute monocytic leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for MIF at approximately 12 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat MIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-289-PB) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

THP-1 cells were stained with Goat Anti-Human/Mouse/Rat MIF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-289-PB, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, CyTOF-ready, Flow
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

MIF Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human MIF
  Pro2-Ala115
  Accession # AAA36315
• Specificity: Detects human, mouse, and rat MIF in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: MIF
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Simple Western 10 ug/mL
  • Western Blot 0.5 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 °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.
• 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
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 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 [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 non-classically 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.

Publications for MIF Antibody (AF-289-PB)(16)

Publications using AF-289-PB

• Michael P. Jeffrey, Chad W. MacPherson, Thomas A. Tompkins, Julia M. Green-Johnson Lacticaseibacillus rhamnosus R0011 secretome attenuates Salmonella enterica serovar Typhimurium secretome-induced intestinal epithelial cell monolayer damage and pro-inflammatory mediator production in intestinal epithelial cell and antigen-presenting cell co-cultures Frontiers in Microbiology 2022-09-28 [PMID: 36246229]
• L Jia, S Rafiei, B Gui, J Wu, XS Liu, AS Kibel Targeting the MIF/CXCR7/AKT Signaling Pathway in Castration-resistant Prostate Cancer Mol. Cancer Res., 2018-09-17;0(0):. 2018-09-17 [PMID: 30224544] (Western Blot, Human)
• J Lang, Y Cheng, A Rolfe, C Hammack, D Vera, K Kyle, J Wang, TB Meissner, Y Ren, C Cowan, H Tang An hPSC-Derived Tissue-Resident Macrophage Model Reveals Differential Responses of Macrophages to ZIKV and DENV Infection Stem Cell Reports, 2018-07-05;0(0):. 2018-07-05 [PMID: 29983385] (Neutralization, Human)
• R Jalili, J Horecka, JR Swartz, RW Davis, HHJ Persson Streamlined circular proximity ligation assay provides high stringency and compatibility with low-affinity antibodies Proc. Natl. Acad. Sci. U.S.A., 2018-01-16;0(0):. 2018-01-16 [PMID: 29339495] (Affinity Assay)
• DiCosmo-Ponticello C, Hoover D, Coffman F, Cohen S, Cohen M MIF inhibits monocytic movement through a non-canonical receptor and disruption of temporal Rho GTPase activities in U-937 cells. Cytokine, 2014-06-06;69(1):47-55. 2014-06-06 [PMID: 25022961] (Western Blot, Human)
• Castriconi R, Dondero A, Bellora F, Moretta L, Castellano A, Locatelli F, Corrias M, Moretta A, Bottino C Neuroblastoma-derived TGF-beta1 modulates the chemokine receptor repertoire of human resting NK cells. J Immunol, 2013-04-10;190(10):5321-8. 2013-04-10 [PMID: 23576682] (Neutralization, Human)
• Fonsato V, Collino F, Herrera M, Cavallari C, Deregibus M, Cisterna B, Bruno S, Romagnoli R, Salizzoni M, Tetta C, Camussi G Human liver stem cell-derived microvesicles inhibit hepatoma growth in SCID mice by delivering antitumor microRNAs. Stem Cells, 2012-09-01;30(9):1985-98. 2012-09-01 [PMID: 22736596] (IHC-P, Human)
• Herrero LJ, Nelson M, Srikiatkhachorn A, Gu R, Anantapreecha S, Fingerle-Rowson G, Bucala R, Morand E, Santos LL, Mahalingam S Critical role for macrophage migration inhibitory factor (MIF) in Ross River virus-induced arthritis and myositis. Proc. Natl. Acad. Sci. U.S.A., 2011-07-05;108(29):12048-53. 2011-07-05 [PMID: 21730129] (IHC-P, Human)
• de Oliveira Gomes A, de Oliveira Silva DA, Silva NM, de Freitas Barbosa B, Franco PS, Angeloni MB, Fermino ML, Roque-Barreira MC, Bechi N, Paulesu LR, Dos Santos MC, Mineo JR, Ferro EA Effect of macrophage migration inhibitory factor (MIF) in human placental explants infected with Toxoplasma gondii depends on gestational age. Am. J. Pathol., 2011-06-01;178(6):2792-801. 2011-06-01 [PMID: 21641401] (IHC-P, Human)
• Pedro L. Vera, Terra E. Wolfe, Alexander E. Braley, Katherine L. Meyer-Siegler Thrombin Induces Macrophage Migration Inhibitory Factor Release and Upregulation in Urothelium: A Possible Contribution to Bladder Inflammation PLoS ONE 2010-12-31 [PMID: 21209875]
• Ansa-Addo EA, Lange S, Stratton D, Antwi-Baffour S, Cestari I, Ramirez MI, McCrossan MV, Inal JM Human plasma membrane-derived vesicles halt proliferation and induce differentiation of THP-1 acute monocytic leukemia cells. J. Immunol., 2010-10-04;185(9):5236-46. 2010-10-04 [PMID: 20921526] (Flow Cytometry, Human)
• Meyer-Siegler KL, Cox J, Leng L, Bucala R, Vera PL Macrophage migration inhibitory factor anti-thrombin III complexes are decreased in bladder cancer patient serum: Complex formation as a mechanism of inactivation. Cancer Lett., 2009-09-16;290(1):49-57. 2009-09-16 [PMID: 19762145] (Western Blot, Human)
• Gilliver SC, Ruckshanthi JP, Hardman MJ, Nakayama T, Ashcroft GS Sex dimorphism in wound healing: the roles of sex steroids and macrophage migration inhibitory factor. Endocrinology, 2008-07-24;149(11):5747-57. 2008-07-24 [PMID: 18653719] (IHC-P, Western Blot, Mouse)
• Daryadel A, Grifone RF, Simon HU, Yousefi S Apoptotic neutrophils release macrophage migration inhibitory factor upon stimulation with tumor necrosis factor-alpha. J. Biol. Chem., 2006-07-21;281(37):27653-61. 2006-07-21 [PMID: 16861224] (ICC, Western Blot, Human)
• Paulesu L, Cateni C, Romagnoli R, Ietta F, Dantzer V Variation in macrophage-migration-inhibitory-factor immunoreactivity during porcine gestation. Biol. Reprod., 2004-11-24;72(4):949-53. 2004-11-24 [PMID: 15564603] (IHC-P, Western Blot, Human, Porcine)
• Chuang CC, Hung CJ, Tsai MC, Chuang YC High concentrations of circulating macrophage migration inhibitory factor in patients with severe blunt trauma: Is serum macrophage migration inhibitory factor concentration a valuable prognostic factor? Crit. Care Med., 2004-03-01;32(3):734-9. 2004-03-01 [PMID: 15090955] (ELISA Development, Human)

Bioinformatics

• Gene Symbol: MIF
• Uniprot:
  • AAA36315()