Purified monocytes (1 × 105) were resuspended in 200 μl of RPMI 1640 media supplemented with 10% FCS, penicillin, and streptomycin. Monocytes were treated for 30 min with the MIF antagonist 3-(3-hydroxybenzyl)-5-methylbenzooxazol-2-one, designated MIF098 (20 μM, at a dose determined by a dose-kinetic study, not shown) (32 (link)) followed by stimulation for 3, 7, or 18 hours with or without U1-snRNP (5 μg/ml, AroTec Diagnostics Limited, New Zealand) in the presence or absence of anti-U1-snRNP Ab+ or healthy serum (final concentration of 5 %) (25 (link)). Some cells were treated for 18 hours with U1-snRNP and anti-U1-snRNP Ab+ serum in the presence or absence of recombinant human MIF (40 μg/ml, R&D Systems, Minneapolis, MN)
Recombinant human mif
Manufactured by R&D Systems
Sourced in United Kingdom
Recombinant human MIF is a protein produced in a laboratory setting. It is a member of the macrophage migration inhibitory factor (MIF) family of proteins. MIF plays a role in the immune response and inflammatory processes.
Automatically generated - may contain errors
2 protocols using recombinant human mif
1
Monocyte Response to U1-snRNP and Anti-U1 Antibodies
2
Characterizing Hematopoietic Cell Phenotypes
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Anti-HIF1α antibody was purchased from BD Biosciences (Oxford, UK). Anti-HIF2α was purchased from (Novus Bio, Cambridge, UK Cat: NB100-122SS) IgG-FITC, IgG-PE, IgG-APC, anti-CD34-PE, anti-CD33-APC and anti-CD45-FITC antibodies were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany Cat. 130098847, 130098845, 130092214, 130098139, 130098043, 130098864). Recombinant human MIF, and MIF ELISA were purchased from R&D Systems (Abingdon, UK). All other reagents were obtained from Sigma-Aldrich (St Louis, MO, USA), unless otherwise indicated
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