Human/Mouse MIF Antibody

(3 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human MIF in Western blots. Because this antibody preparation is a total IgG fration, complete monospecificity cannot be assumed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Protein A or G purified
  • Immunogen
    E. coli-derived recombinant human MIF (Catalog # 289-MF)
    Pro2-Ala115
    Accession # AAA36315
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2-1 µg/mL
    See below
  • Simple Western
    25 µg/mL
    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.
Data Examples
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 0.2 µg/mL of Goat Anti-Human/Mouse MIF Polyclonal Antibody (Catalog # AB-289-PB) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # 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 Human MIF by Simple WesternTM. Simple Western lane view shows lysates of THP‑1 human acute monocytic leukemia cell line and U937 human histiocytic lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for MIF at approximately 12 kDa (as indicated) using 25 µg/mL of Goat Anti-Human/Mouse MIF Polyclonal Antibody (Catalog #
AB-289-PB) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Mouse MIF by Western Blot. Western blot shows lysates of J774A.1 mouse reticulum cell sarcoma macrophage cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse MIF Polyclonal Antibody (Catalog # AB-289-PB) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # 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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 1 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it 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:
    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
Related Research Areas
Citations:

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.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. UVA induces granzyme B in human keratinocytes through MIF: implication in extracellular matrix remodeling.
    Authors: Hernandez-Pigeon H, Jean C, Charruyer A, Haure MJ, Baudouin C, Charveron M, Quillet-Mary A, Laurent G
    J. Biol. Chem., 2007;282(11):8157-64.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  2. The sex steroid precursor DHEA accelerates cutaneous wound healing via the estrogen receptors.
    Authors: Mills SJ, Ashworth JJ, Gilliver SC, Hardman MJ, Ashcroft GS
    J. Invest. Dermatol., 2005;125(5):1053-62.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  3. Estrogen modulates cutaneous wound healing by downregulating macrophage migration inhibitory factor.
    Authors: Ashcroft GS, Mills SJ, Lei K, Gibbons L, Jeong MJ, Taniguchi M, Burow M, Horan MA, Wahl SM, Nakayama T
    J. Clin. Invest., 2003;111(9):1309-18.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
Isotype Controls
Description Application Cat# Citations Images  

Normal Goat IgG Control

Ctrl AB-108-C 191  
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Secondary Antibodies
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Goat IgG HRP-conjugated Antibody

WB, Simple Western HAF109 19  
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Goat IgG HRP-conjugated Antibody

WB HAF017 15  
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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 9
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 6  
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Donkey Anti-Goat IgG Antibody

WB AF109 6
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Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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