Mouse Macrophage Migration Inhibitory Factor (MIF) ELISA Kit

Principle of the Assay

The microtiter plate provided in this kit has been pre-coated with an antibody specific to MIF. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to MIF. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After the TMB substrate solution is added, only those wells that contain MIF, biotin-conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution, and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of MIF in the samples is then determined by comparing the O.D. of the samples to the standard curve.

For Use with serum, plasma, and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.

Target Information

Pro-inflammatory cytokine involved in the innate immune response to bacterial pathogens (By similarity). The expression of MIF at sites of inflammation suggests a role as mediator in regulating the function of macrophages in host defense (By similarity). Counteracts the anti-inflammatory activity of glucocorticoids (By similarity). Has phenylpyruvate tautomerase and dopachrome tautomerase activity (in vitro), but the physiological substrate is not known (PubMed:19188446, PubMed:10933783, PubMed:16780921). It is not clear whether the tautomerase activity has any physiological relevance, and whether it is important for cytokine activit (PubMed:19188446, PubMed:10933783, PubMed:16780921).

GENE ID	17319
SWISS PROT	P34884
SYNONYMS	GIF; GLIF; MMIF; Glycosylation-Inhibiting Factor; L-dopachrome isomerase; L-dopachrome tautomerase; Phenylpyruvate tautomerase

Materials Supplied

Kit Components	96 Wells Quantity/Size
Pre-coated, ready-to-use 96-well strip plate	1 plate
Plate sealer for 96 wells	2
Standard	2 tubes
Diluent buffer	1 bottle
Detection Reagent A	1 bottle
Detection Reagent B	1 bottle
TMB Substrate	1 tube
Stop Solution	1 tube
Wash Buffer (30 ℅ concentrate)	1 tube
Product data sheet	1 copy

Storage

The TMB Substrate, Wash Buffer (30X concentrate), and the Stop Solution should be stored at 4°C upon receipt, while the other items should be stored at -20°C.

Performance Characteristics

REPEATABILITY	Intra-assay Precision (Precision within an assay): 3 samples with low, middle, and high-level MIF were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle, and high-level MIF were tested on 3 different plates, with 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
SENSITIVITY	The minimum detectable dose was 13.3pg/mL.
ASSAY RANGE	31.2-2000pg/mL
SPECIFICITY	This assay has high sensitivity and excellent specificity for the detection of MIF. No significant cross-reactivity or interference between MIF and analogs was observed. Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between MIF and all analogs, therefore, cross reactivity may still exist.