Human MIF Alexa Fluor® 750-conjugated Antibody

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IC2891S-100UG

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Human MIF Alexa Fluor® 750-conjugated Antibody Summary

Species Reactivity
Human
Specificity
Detects human MIF in direct ELISAs.
Source
Monoclonal Mouse IgG1 Clone # 932606
Immunogen
E. coli-derived recombinant human MIF
Met1-Phe114
Accession # P14174
Formulation
Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 750 (Excitation= 749 nm, Emission= 775 nm)

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
0.25-1 µg/106 cells
Human peripheral blood mononuclear cell (PBMCs) treated with LPS overnight and monensin for 2 hours were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

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.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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

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Product Specific Notices


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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