Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Bovine, Transgenic Mouse

Applications

Validated:

Western Blot, Simple Western

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Immunocytochemistry, Immunoprecipitation, ELISA Capture, ELISA Development, ELISA Development (Capture), Luminex Development

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 12302
View all MIF Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human MIF
Pro2-Ala115
Accession # P14174

Specificity

Detects human MIF in Western blots. In Western blots, this antibody shows approximately 25% cross-reactivity with recombinant mouse MIF.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human MIF Antibody

Detection of Human MIF antibody by Western Blot.

Detection of Human MIF by Western Blot.

Western blot shows lysate of K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human MIF Monoclonal Antibody (Catalog # MAB289) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 Exosome Standards (HEK293) (NBP3-11684), Exosome Standards (LNCaP) (NBP3-11687) and K562 human chronic myelogenous leukemia cell line, loaded at 0.5 mg/ml. A specific band was detected for MIF at approximately 11 kDa (as indicated) using 10 µg/ml of Mouse Anti-Human MIF Monoclonal Antibody (Catalog # MAB289) followed by HRP-conjugated Goat Anti-Mouse Secondary Antibody (Catalog # 042-205). This experiment was conducted under reducing conditions and using the 2-40kDa separation system.
Detection of Humean MIF antibody by Simple WesternTM.

Detection of Humean 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 10 µg/mL of Mouse Anti-Human MIF Monoclonal Antibody (Catalog # MAB289). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of MIF by Western Blot

Detection of MIF by Western Blot

Effect of treatments on MIF secretion, intracellular content and gene expression. A. MIF secretion – Conditioned medium following 24 h growth was assayed for MIF content by ELISA. Data are expressed as culture medium MIF concentration in pg per 105 cells. All treatments decreased MIF in the culture media. B. Western blot of anti-MIF treated conditioned medium following 24 h treatment- 12 kDa MIF band is present in all samples. Lane 1, DMEM 1% BSA; Lane 2, non-specific mouse IgG1; Lane 3, anti-MIF antibody treatment. C. Intracellular MIF content – Changes in intracellular MIF 24 h post treatment. Data are expressed as cleared cell lysate MIF concentration in pg per 105 cells. Note that HA treatment increased, while MIF antisense treatment decrease the concentration of MIF in the cell lysates. Anti-MIF treatment did not produce a significant effect. D. MIF gene expression -MIF mRNA content was quantified in cells following 24 h and the indicated treatment. Data are expressed as a relative intensity ratio. PCR band intensity was determined from the formula, total intensity = area × average intensity. The relative intensity ratio is determined from the total intensity of gene specific PCR product band divided by the 18 S rRNA band intensity (internal standard). All treatments resulted in a decrease in MIF mRNA with the greatest effects seen following MIF anti-sense treatment. (* – p < 0.05, ** – p < 0.01, *** – p < 0.001). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/15248897), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human MIF Antibody

Application
Recommended Usage

Simple Western

10 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line and U937 human histiocytic lymphoma cell line

Western Blot

1 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line

Reviewed Applications

Read 8 reviews rated 4.4 using MAB289 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

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

P14174

Additional MIF Products

Product Documents for Human MIF Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human MIF Antibody

For research use only

Citations for Human MIF Antibody

Customer Reviews for Human MIF Antibody (8)

4.4 out of 5
8 Customer Ratings
5 Stars
50%
4 Stars
38%
3 Stars
13%
2 Stars
0%
1 Stars
0%

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Customer Images

Showing  1 - 5 of 8 reviews Showing All
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  • Human MIF Antibody
    Name: Katie Myers
    Application: ELISA
    Sample Tested: Serum
    Species: Primate
    Verified Customer | Posted 11/14/2022
    Used as a non-human primate capture molecule
    Human MIF Antibody MAB289
  • Human CD300e/LMIR6 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: THP-1 human acute monocytic leukemia cell line
    Species: Human
    Verified Customer | Posted 10/07/2021
    Western blot. THP-1 human acute monocytic leukemia cell line.
    Human MIF Antibody MAB289
  • Human MIF Antibody
    Name: Anonymous
    Application: Microarrays
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 01/14/2021
    Human MIF Antibody MAB289
  • Human MIF Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 07/19/2019
    Human MIF Antibody MAB289
  • Human MIF Antibody
    Name: Carly Garrison
    Application: Microarray
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 11/20/2018
  • Human MIF Antibody
    Name: Anonymous
    Application: Microarrays
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 11/14/2018
  • Human MIF Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 11/08/2018
  • Human MIF Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 11/07/2017
    This antibody was used as a matched pair with AF289 to build a sandwich ELISA to measure MIF in human serum and plasma. The antibody worked well.
    Human MIF Antibody MAB289

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