Human MIF DuoSet ELISA

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Human MIF ELISA Standard Curve
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Product Details
Citations (32)
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Reviews (3)

Human MIF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human MIF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.


Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Human MIF ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. 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 (macrophage migration inhibitory factor) is a pro-inflammatory cytokine that is released during microbial infection, ARDS, rheumatoid arthritis, atherosclerosis, type 2 diabetes, and cancer. It signals through CXCR4, CXCR7, or complexes of CD74-CD44. MIF exhibits potent paracrine and autocrine stimulatory effects on cell growth and survival and enhances the production of inflammatory cytokines. It also inhibits the effectiveness of immunosuppressive glucocorticoids.

Long Name:
Macrophage Migration Inhibitory Factor
Entrez Gene IDs:
4282 (Human); 17319 (Mouse)
Alternate Names:
EC; EC; 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

Assay Procedure


Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.


Citations for Human MIF DuoSet ELISA

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.

32 Citations: Showing 1 - 10
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  1. Distinct patterns of serum and urine macrophage migration inhibitory factor kinetics predict death in sepsis: a prospective, observational clinical study
    Authors: J Toldi, L Kelava, S Marton, D Muhl, P Kustan, Z Feher, K Maar, J Garai, E Pakai, A Garami
    Scientific Reports, 2023-01-11;13(1):588.
    Species: Human
    Sample Types: Serum
  2. Macrophage Migration Inhibitory Factor in Major Depressive Disorder: A Multilevel Pilot Study
    Authors: C Swoboda, L Deloch, C von Zimmer, T Richter-Sc, B Lenz, J Kornhuber, C Mühle
    International Journal of Molecular Sciences, 2022-12-07;23(24):.
    Species: Human
    Sample Types: Serum
  3. Evaluation of Immunomodulatory Responses and Changed Wound Healing in Type 2 Diabetes-A Study Exploiting Dermal Fibroblasts from Diabetic and Non-Diabetic Human Donors
    Authors: K Nickel, U Wensorra, H Wenck, N Peters, H Genth
    Cells, 2021-10-28;10(11):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Biomarkers of disease severity in patients with visceral leishmaniasis co-infected with HIV
    Authors: GR Ferreira, JR Santos-Oli, ML Silva-Frei, M Honda, DL Costa, AM Da-Cruz, CHN Costa
    Cytokine, 2021-10-27;149(0):155747.
    Species: Human
    Sample Types: Serum
  5. Heme Oxygenase-1 Induction in Human BeWo Trophoblast Cells Decreases Toxoplasma gondii Proliferation in Association With the Upregulation of p38 MAPK Phosphorylation and IL-6 Production
    Authors: MPO Almeida, CM Mota, TWP Mineo, EAV Ferro, BF Barbosa, NM Silva
    Frontiers in Microbiology, 2021-04-12;12(0):659028.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Associations between salivary cytokines and periodontal and microbiological parameters in orthodontic patients
    Authors: Y Chen, WK Wong, JC Seneviratn, S Huang, C McGrath, U Hagg
    Medicine, 2021-03-12;100(10):e24924.
    Species: Human
    Sample Types: Saliva
  7. Macrophage migration inhibitory factor may play a protective role in osteoarthritis
    Authors: M Liu, Z Xie, G Sun, L Chen, D Qi, H Zhang, J Xiong, A Furey, P Rahman, G Lei, G Zhai
    Arthritis Research & Therapy, 2021-02-20;23(1):59.
    Species: Human
    Sample Types: Plasma
  8. Feedback activation of STAT3 limits the response to PI3K/AKT/mTOR inhibitors in PTEN-deficient cancer cells
    Authors: J Wang, X Lv, X Guo, Y Dong, P Peng, F Huang, P Wang, H Zhang, J Zhou, Y Wang, B Wei, ZF Shang, S Li
    Oncogenesis, 2021-01-05;10(1):8.
  9. Copaifera spp. oleoresins impair Toxoplasma gondii infection in both human trophoblastic cells and human placental explants
    Authors: SC Teixeira, G de Souza, BC Borges, TE de Araújo, AM Rosini, FA Aguila, SR Ambrósio, RCS Veneziani, JK Bastos, MJB Silva, CHG Martins, B de Freitas, EAV Ferro
    Scientific Reports, 2020-09-16;10(1):15158.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Brazilian strains of Toxoplasma gondii are controlled by azithromycin and modulate cytokine production in human placental explants
    Authors: PS Franco, PSG Gois, TE de Araújo, RJ da Silva, B de Freitas, A de Oliveir, F Ietta, LA Dos Santos, MC Dos Santos, JR Mineo, EAV Ferro
    J. Biomed. Sci., 2019-01-21;26(1):10.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Involvement of HPV Infection in the Release of Macrophage Migration Inhibitory Factor in Head and Neck Squamous Cell Carcinoma
    Authors: N Kindt, G Descamps, JR Lechien, M Remmelink, JM Colet, R Wattiez, G Berchem, F Journe, S Saussez
    J Clin Med, 2019-01-10;8(1):.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Silver nanoparticles promote the emergence of heterogeneic human neutrophil sub-populations
    Authors: JA Fraser, S Kemp, L Young, M Ross, M Prach, GR Hutchison, E Malone
    Sci Rep, 2018-05-14;8(1):7506.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Caspase-8 Acts in a Non-enzymatic Role as a Scaffold for Assembly of a Pro-inflammatory FADDosome Complex upon TRAIL Stimulation
    Authors: CM Henry, SJ Martin
    Mol. Cell, 2017-02-16;65(4):715-729.e5.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Secretory activity of subcutaneous abdominal adipose tissue in male patients with rheumatoid arthritis and osteoarthritis - association with clinical and laboratory data
    Authors: Ewa Kontny
    Reumatologia, 2016-11-28;54(5):227-235.
    Species: Human
    Sample Types: Tissue Explants
  15. The development of a novel Mycobacterium-Escherichia coli shuttle vector system using pMyong2, a linear plasmid from Mycobacterium yongonense DSM 45126T.
    Authors: Lee H, Kim B, Kim B, Kook Y, Kim B
    PLoS ONE, 2015-03-30;30(0):.
    Species: Bacteria - Mycobacterium smegmatis
    Sample Types: Cell Lysates
  16. Involvement of macrophage migration inhibitory factor and its receptor (CD74) in human breast cancer.
    Authors: Richard V, Kindt N, Decaestecker C, Gabius H, Laurent G, Noel J, Saussez S
    Oncol Rep, 2014-06-16;32(2):523-9.
    Species: Human
    Sample Types: Serum
  17. Rescue of impaired NK cell activity in hodgkin lymphoma with bispecific antibodies in vitro and in patients.
    Authors: Reiners K, Kessler J, Sauer M, Rothe A, Hansen H, Reusch U, Hucke C, Kohl U, Durkop H, Engert A, von Strandmann E
    Mol Ther, 2013-03-05;21(4):895-903.
    Species: Human
    Sample Types: Serum
  18. Elevated serum levels of macrophage migration inhibitory factor are associated with progressive chronic cardiomyopathy in patients with Chagas disease.
    Authors: Cutrullis R, Petray P, Schapachnik E, Sanchez R, Postan M, Gonzalez M, Martin V, Corral R
    PLoS ONE, 2013-02-22;8(2):e57181.
    Species: Human
    Sample Types: Serum
  19. Evidence that lipopolisaccharide may contribute to the cytokine storm and cellular activation in patients with visceral leishmaniasis.
    Authors: Santos-Oliveira JR, Regis EG, Leal CR, Cunha RV, Bozza PT, Da-Cruz AM
    PLoS Negl Trop Dis, 2011-07-12;5(7):e1198.
    Species: Human
    Sample Types: Plasma
  20. 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: Cell Culture Supernates
  21. 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
  22. A fluorinated analog of ISO-1 blocks the recognition and biological function of MIF and is orally efficacious in a murine model of colitis.
    Authors: Dagia NM, Kamath DV, Bhatt P, Gupte RD, Dadarkar SS, Fonseca L, Agarwal G, Chetrapal-Kunwar A, Balachandran S, Srinivasan S, Bose J, Pari K, B-Rao C, Parkale SS, Gadekar PK, Rodge AH, Mandrekar N, Vishwakarma RA, Sharma S
    Eur. J. Pharmacol., 2009-02-21;607(1):201-12.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Up-regulated macrophage migration inhibitory factor protects apoptosis of dermal fibroblasts in patients with systemic sclerosis.
    Authors: Kim JY, Kwok SK, Hur KH, Kim HJ, Kim NS, Yoo SA, Kim WU, Cho CS
    Clin. Exp. Immunol., 2008-03-18;152(2):328-35.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Suspension microarrays for the identification of the response patterns in hyperinflammatory diseases.
    Authors: Hsu HY, Wittemann S, Schneider EM, Weiss M, Joos TO
    Med Eng Phys, 2008-03-03;30(8):976-83.
    Species: Human
    Sample Types: Plasma
  25. The effects of dexamethasone and dehydroepiandrosterone (DHEA) on cytokines and receptor expression in a human osteoblastic cell line: potential steroid-sparing role for DHEA.
    Authors: Harding G, Mak YT, Evans B, Cheung J, MacDonald D, Hampson G
    Cytokine, 2006-12-11;36(1):57-68.
    Species: Human
    Sample Types: Cell Culture Supernates
  26. Exogenous platelet-activating factor acetylhydrolase reduces mortality in mice with systemic inflammatory response syndrome and sepsis.
    Authors: Gomes RN, Bozza FA, Amancio RT, Japiassu AM, Vianna RC, Larangeira AP, Gouvea JM, Bastos MS, Zimmerman GA, Stafforini DM, Prescott SM, Bozza PT, Castro-Faria-Neto HC
    Shock, 2006-07-01;26(1):41-9.
    Species: Mouse
    Sample Types: Peritoneal Fluid
  27. Glucocorticoids suppress macrophage migration inhibitory factor (MIF) expression in a cell-type-specific manner.
    Authors: Alourfi Z, Donn RP, Stevens A, Berry A, McMaster A, Ray DW
    J. Mol. Endocrinol., 2005-04-01;34(2):583-95.
    Species: Human
    Sample Types: Cell Culture Supernates
  28. A functional promoter haplotype of macrophage migration inhibitory factor is linked and associated with juvenile idiopathic arthritis.
    Authors: Donn R, Alourfi Z, Zeggini E, Lamb R, Jury F, Lunt M, Meazza C, De Benedetti F, Thomson W, Ray D, Ray D
    Arthritis Rheum., 2004-05-01;50(5):1604-10.
    Species: Human
    Sample Types: Plasma
  29. Reduced levels of transforming growth factor-beta1, interleukin-12 and increased migration inhibitory factor are associated with severe malaria.
    Authors: Chaiyaroj SC, Rutta AS, Muenthaisong K, Watkins P, Na Ubol M, Looareesuwan S
    Acta Trop., 2004-02-01;89(3):319-27.
    Species: Human
    Sample Types: Plasma
  30. Interleukin-8 serum levels in patients with hepatocellular carcinoma: correlations with clinicopathological features and prognosis.
    Authors: Ren Y, Poon RT, Tsui HT, Chen WH, Li Z, Lau C, Yu WC, Fan ST
    Clin. Cancer Res., 2003-12-01;9(16):5996-6001.
    Species: Human
    Sample Types: Serum
  31. The cytokine macrophage migration inhibitory factor reduces pro-oxidative stress-induced apoptosis.
    Authors: Nguyen MT, Lue H, Kleemann R, Thiele M, Tolle G, Finkelmeier D, Wagner E, Braun A, Bernhagen J
    J. Immunol., 2003-03-15;170(6):3337-47.
    Species: Human
    Sample Types: Cell Lysates
  32. Urine macrophage migration inhibitory factor reflects the severity of renal injury in human glomerulonephritis.
    Authors: Brown FG, Nikolic-Paterson DJ, Hill PA, Isbel NM, Dowling J, Metz CM, Atkins RC
    J. Am. Soc. Nephrol., 2002-01-01;13(0):S7-13.
    Species: Human
    Sample Types: Serum


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Human MIF DuoSet ELISA
By Anonymous on 11/01/2022
Sample Tested: iDC human immature myeloid dendritic cells

We used two batch lot number of this kit. Both work good.

Human MIF DuoSet ELISA
By Anonymous on 10/04/2022
Sample Tested: iDC human immature myeloid dendritic cells

Human MIF DuoSet ELISA
By Anonymous on 07/06/2021
Sample Tested: Serum and Plasma

we used this kit to quantify human MIF in serum and plasma samples. Works very well and easy to use protocol.