Human MIF ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Macrophage migration inhibitory factor (MIF) is a protein which in humans is encoded by the MIF gene. This gene is located to human chromosome 22q11.2.3 It is remarkably small; it has 3 exons separated by introns of only 189 and 95 bp, and covers less than 1 kb.4 This gene encodes a lymphokine that may be involved in cell-mediated immunity, immunoregulation, and inflammation.5 MIF plays a role in the regulation of macrophage function in host defense through the suppression of anti-inflammatory effects of glucocorticoids. This lymphokine and the JAB1 protein might form a complex in the cytosol near the peripheral plasma membrane, which may possibly indicate a role in integrin signaling pathways. MIF also plays a central role in the toxic response to endotoxemia and possibly septic shock.Macrophage migration inhibitory factor has been reported to interact with COP9 constitutive photomorphogenic homolog subunit 5, CD74, BNIPL, and CXCR4.

Macrophage migration inhibitory factor; MIF; Glycosylation-inhibiting factor; GIF; L-dopachrome isomerase; L-dopachrome tautomerase; Phenylpyruvate tautomerase; MIF; GLIF; MMIF

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 1442 3826 6427 1625 4177 6782 Standard deviation 90.85 206.6 315 121.9 254.8 393.4 CV (%) 6.3% 5.4% 4.9% 7.5% 6.1% 5.8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 1442 3826 6427 1625 4177 6782 Standard deviation 90.85 206.6 315 121.9 254.8 393.4 CV (%) 6.3% 5.4% 4.9% 7.5% 6.1% 5.8% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 1442 3826 6427 1625 4177 6782 Mean (pg/ml) 1442 3826 6427 1625 4177 6782 Standard deviation 90.85 206.6 315 121.9 254.8 393.4 Standard deviation 90.85 206.6 315 121.9 254.8 393.4 CV (%) 6.3% 5.4% 4.9% 7.5% 6.1% 5.8% CV (%) 6.3% 5.4% 4.9% 7.5% 6.1% 5.8%

Pro-inflammatory cytokine. Involved in the innate immune response to bacterial pathogens. The expression of MIF at sites of inflammation suggests a role as mediator in regulating the function of macrophages in host defense. Counteracts the anti- inflammatory activity of glucocorticoids. Has phenylpyruvate tautomerase and dopachrome tautomerase activity (in vitro), but the physiological substrate is not known. It is not clear whether the tautomerase activity has any physiological relevance, and whether it is important for cytokine activity.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0813-CA Rabbit anti- human MIF polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0813-DA Biotinylated goat anti- human MIF polyclonal antibody (Detection Antibody) 500 μl, 14 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0813-ST Lyophilized recombinant human MIF standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0813-CA Rabbit anti- human MIF polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0813-DA Biotinylated goat anti- human MIF polyclonal antibody (Detection Antibody) 500 μl, 14 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0813-ST Lyophilized recombinant human MIF standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0813-CA Rabbit anti- human MIF polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0813-CA Rabbit anti- human MIF polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0813-DA Biotinylated goat anti- human MIF polyclonal antibody (Detection Antibody) 500 μl, 14 μg/mL (recommended dilution 1:100) EZ0813-DA Biotinylated goat anti- human MIF polyclonal antibody (Detection Antibody) 500 μl, 14 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0813-ST Lyophilized recombinant human MIF standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0813-ST Lyophilized recombinant human MIF standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.046 0.111 0.215 0.357 0.635 1.063 1.582 1.999 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.046 0.111 0.215 0.357 0.635 1.063 1.582 1.999 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.046 0.111 0.215 0.357 0.635 1.063 1.582 1.999 O.D. 0.046 0.111 0.215 0.357 0.635 1.063 1.582 1.999

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)