Human/Mouse/Rat MIF Antibody Summary
Pro2-Ala115
Accession # AAA36315
Applications
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
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 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.
Detection of Mouse and Rat MIF by Western Blot. 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.
Detection of Human MIF by Simple WesternTM. 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.
Detection of MIF in THP-1 cells by Flow Cytometry. 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.
Preparation and Storage
- 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 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.
- 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.
Product Datasheets
Citations for Human/Mouse/Rat MIF Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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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
Authors: Michael P. Jeffrey, Chad W. MacPherson, Thomas A. Tompkins, Julia M. Green-Johnson
Frontiers in Microbiology
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Targeting the MIF/CXCR7/AKT Signaling Pathway in Castration-resistant Prostate Cancer
Authors: L Jia, S Rafiei, B Gui, J Wu, XS Liu, AS Kibel
Mol. Cancer Res., 2018-09-17;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
An hPSC-Derived Tissue-Resident Macrophage Model Reveals Differential Responses of Macrophages to ZIKV and DENV Infection
Authors: J Lang, Y Cheng, A Rolfe, C Hammack, D Vera, K Kyle, J Wang, TB Meissner, Y Ren, C Cowan, H Tang
Stem Cell Reports, 2018-07-05;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization -
Streamlined circular proximity ligation assay provides high stringency and compatibility with low-affinity antibodies
Authors: R Jalili, J Horecka, JR Swartz, RW Davis, HHJ Persson
Proc. Natl. Acad. Sci. U.S.A., 2018-01-16;0(0):.
Applications: Affinity Assay -
MIF inhibits monocytic movement through a non-canonical receptor and disruption of temporal Rho GTPase activities in U-937 cells.
Authors: DiCosmo-Ponticello C, Hoover D, Coffman F, Cohen S, Cohen M
Cytokine, 2014-06-06;69(1):47-55.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Neuroblastoma-derived TGF-beta1 modulates the chemokine receptor repertoire of human resting NK cells.
Authors: Castriconi R, Dondero A, Bellora F, Moretta L, Castellano A, Locatelli F, Corrias M, Moretta A, Bottino C
J Immunol, 2013-04-10;190(10):5321-8.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization -
Human liver stem cell-derived microvesicles inhibit hepatoma growth in SCID mice by delivering antitumor microRNAs.
Authors: Fonsato V, Collino F, Herrera M, Cavallari C, Deregibus M, Cisterna B, Bruno S, Romagnoli R, Salizzoni M, Tetta C, Camussi G
Stem Cells, 2012-09-01;30(9):1985-98.
Species: Human
Sample Types: Whole Cells
Applications: IHC-P -
Critical role for macrophage migration inhibitory factor (MIF) in Ross River virus-induced arthritis and myositis.
Authors: Herrero LJ, Nelson M, Srikiatkhachorn A, Gu R, Anantapreecha S, Fingerle-Rowson G, Bucala R, Morand E, Santos LL, Mahalingam S
Proc. Natl. Acad. Sci. U.S.A., 2011-07-05;108(29):12048-53.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Effect of macrophage migration inhibitory factor (MIF) in human placental explants infected with Toxoplasma gondii depends on gestational age.
Authors: 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
Am. J. Pathol., 2011-06-01;178(6):2792-801.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Thrombin Induces Macrophage Migration Inhibitory Factor Release and Upregulation in Urothelium: A Possible Contribution to Bladder Inflammation
Authors: Pedro L. Vera, Terra E. Wolfe, Alexander E. Braley, Katherine L. Meyer-Siegler
PLoS ONE
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Human plasma membrane-derived vesicles halt proliferation and induce differentiation of THP-1 acute monocytic leukemia cells.
Authors: Ansa-Addo EA, Lange S, Stratton D, Antwi-Baffour S, Cestari I, Ramirez MI, McCrossan MV, Inal JM
J. Immunol., 2010-10-04;185(9):5236-46.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Macrophage migration inhibitory factor anti-thrombin III complexes are decreased in bladder cancer patient serum: Complex formation as a mechanism of inactivation.
Authors: Meyer-Siegler KL, Cox J, Leng L, Bucala R, Vera PL
Cancer Lett., 2009-09-16;290(1):49-57.
Species: Human
Sample Types: Serum
Applications: Western Blot -
Sex dimorphism in wound healing: the roles of sex steroids and macrophage migration inhibitory factor.
Authors: Gilliver SC, Ruckshanthi JP, Hardman MJ, Nakayama T, Ashcroft GS
Endocrinology, 2008-07-24;149(11):5747-57.
Species: Mouse
Sample Types: Tissue Homogenates, Whole Tissue
Applications: IHC-P, Western Blot -
Apoptotic neutrophils release macrophage migration inhibitory factor upon stimulation with tumor necrosis factor-alpha.
Authors: Daryadel A, Grifone RF, Simon HU, Yousefi S
J. Biol. Chem., 2006-07-21;281(37):27653-61.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: ICC, Western Blot -
Variation in macrophage-migration-inhibitory-factor immunoreactivity during porcine gestation.
Authors: Paulesu L, Cateni C, Romagnoli R, Ietta F, Dantzer V
Biol. Reprod., 2004-11-24;72(4):949-53.
Species: Human, Porcine
Sample Types: Tissue Homogenates, Whole Tissue
Applications: IHC-P, Western Blot -
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?
Authors: Chuang CC, Hung CJ, Tsai MC, Chuang YC
Crit. Care Med., 2004-03-01;32(3):734-9.
Species: Human
Sample Types: Serum
Applications: ELISA Development
FAQs
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What does the PB in AF-289-PB mean?
The PB in AF-289-PB is a part of the catalog code for this antibody and it appears on the vial lable for both the retail(100 µg) and SP (25 µg) size of the antibody.
Reviews for Human/Mouse/Rat MIF Antibody
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