Human/Mouse/Rat MIF Antibody

R&D Systems, part of Bio-Techne | Catalog # AF-289-PB

R&D Systems, part of Bio-Techne
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AF-289-PB
AF-289-SP
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Conjugate
Catalog #
Biotin
BAF289
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Porcine

Applications

Validated:

CyTOF-ready, Intracellular Staining by Flow Cytometry, Simple Western, Western Blot

Cited:

Affinity Assay, ELISA Development, Flow Cytometry, Immunocytochemistry, Immunohistochemistry-Paraffin, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all MIF Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human MIF
Pro2-Ala115
Accession # AAA36315

Specificity

Detects human, mouse, and rat MIF in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat MIF Antibody

Detection of Human MIF antibody by Western Blot.

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 antibody by Western Blot.

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.
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Applications for Human/Mouse/Rat MIF Antibody

Application
Recommended Usage

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
Sample: THP-1 cells fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

Simple Western

10 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line

Western Blot

0.5 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line, U937 human histiocytic lymphoma cell line, J774A.1 mouse reticulum cell sarcoma macrophage cell line, and NR8383 rat alveolar macrophage cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using AF-289-PB in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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 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

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), MMIF, Phenylpyruvate tautomerase

Entrez Gene IDs

4282 (Human); 17319 (Mouse)

Gene Symbol

MIF

UniProt

AAA36315

Additional MIF Products

Product Documents for Human/Mouse/Rat MIF Antibody

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Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat MIF Antibody

For research use only

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