Human MIF DuoSet ELISA
Human MIF DuoSet ELISA Summary
* 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
Product Datasheets
Preparation and Storage
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.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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.
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Citations: Showing 1 - 10
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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
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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
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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
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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
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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
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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
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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
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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.
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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
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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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