Human/Mouse/Rat MIF Antibody

Catalog # Availability Size / Price Qty
AF-289-PB
AF-289-SP

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Detection of Human MIF by Western Blot.
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Product Details
Citations (16)
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Reviews (2)

Human/Mouse/Rat MIF Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse, and rat MIF in Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human MIF
Pro2-Ala115
Accession # AAA36315
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
0.5 µg/mL
See below
Simple Western
10 µg/mL
THP‑1 human acute monocytic leukemia cell line
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
THP-1 cells fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

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 Detection of Human MIF antibody by Western Blot. View Larger

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.

Western Blot Detection of Mouse and Rat MIF antibody by Western Blot. View Larger

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.

Simple Western View Larger

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.

Intracellular Staining by Flow Cytometry View Larger

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.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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 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.

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
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

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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.

16 Citations: Showing 1 - 10
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  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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

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Human/Mouse/Rat MIF Antibody
By Anonymous on 07/19/2019
Application: ELISA Sample Tested: Serum and Plasma Species: Human

Human/Mouse/Rat MIF Antibody
By Anonymous on 11/07/2017
Application: ELISA Sample Tested: Serum and Plasma Species: Human

This polyclonal was paired up with the monoclonal MAB289 to build a sandwich ELISA for the measurement of human MIF in serum and plasma samples.