Human MIF ELISA Kit EZ-Set™ (DIY Antibody Pairs) (antibody pairs and standards for assay development). Quantitate Human MIF in cell culture supeatants, serum, plasma (heparin, EDTA) and human milk.
Expression system for standard: E.coli; Immunogen sequence: P2-A115
Kit Components
Description Capture Antibody Detection Antibody Avidin-Biotin-Peroxidase Complex (ABC) Lyophilized Standard
Cross Reactivity
There is no detectable cross-reactivity with other relevant proteins.
Specificity
Natural and recombinant human MIF
Subcellular Localization
Secreted.
Sample Type
Cell culture supeatants, Serum, Plasma, Human milk
Anticoagulant
heparin or EDTA
Precision
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%
Protein Function
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.
Background
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.
Notes
Material Required But Not Provided: 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 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 11. Stop Solution: 2 N H2SO4 12. Wash Buffer . 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
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.)
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