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EK000079-EK0813: Human MIF ELISA Kit
Range 156pg/ml-10,000pg/ml
Sensitivity < 20pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitative detection of human MIF in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Macrophage migration inhibitory factor (MIF) is a protein which in humans is encoded by the MIF gene.1, 2 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.6 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.7 Macrophage migration inhibitory factor has been reported to interact with COP9 constitutive photomorphogenic homolog subunit 5, CD74, BNIPL, and CXCR4. The standard product used in this kit is recombinant human MIF, consisting of 115 amino acids with the molecular mass of 12.5kDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Weiser WY, Temple PA, Witek-Giannotti JS, Remold HG, Clark SC, David JR (October 1989). "Molecular cloning of a cDNA encoding a human macrophage migration inhibitory factor". Proc. Natl. Acad. Sci. U.S.A. 86 (19): 7522?.
2. Kozak CA, Adamson MC, Buckler CE, Segovia L, Paralkar V, Wistow G (June 1995). "Genomic cloning of mouse MIF (macrophage inhibitory factor) and genetic mapping of the human and mouse expressed gene and nine mouse pseudogenes". Genomics 27 (3): 405?1.
3. Budarf, M.; McDonald, T.; Sellinger, B.; Kozak, C.; Graham, C.; Wistow, G. : Localization of the human gene for macrophage migration inhibitory factor (MIF) to chromosome 22q11.2. Genomics 39: 235-236, 1997.
4. Paralkar, V.; Wistow, G. : Cloning the human gene for macrophage migration inhibitory factor (MIF). Genomics 19: 48-51, 1994.
5. Leng L, Bucala R (February 2006). "Insight into the biology of macrophage migration inhibitory factor (MIF) revealed by the cloning of its cell surface receptor.". Cell Research 16 (2): 162-8.
6. Flaster H, Bernhagen J, Calandra T, Bucala R (June 2007). "The macrophage migration inhibitory factor-glucocorticoid dyad: regulation of inflammation and immunity.". Mol Endocrinol. 21 (6): 1267-80.
7. Bernhagen, J.; Calandra, T.; Mitchell, R. A.; Martin, S. B.; Tracey, K. J.; Voelter, W.; Manogue, K. R.; Cerami, A.; Bucala, R. : MIF is a pituitary-derived cytokine that potentiates lethal endotoxaemia. Nature 365: 756-759, 1993.
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