Multiplex Assay Kit for Macrophage Migration Inhibitory Factor (MIF) ,etc. by FLIA (Flow Luminescence Immunoassay)

GIF; GLIF; MMIF; Glycosylation-Inhibiting Factor; L-dopachrome isomerase; L-dopachrome tautomerase; Phenylpyruvate tautomerase

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Macrophage Migration Inhibitory Factor (MIF) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Macrophage Migration Inhibitory Factor (MIF) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Macrophage Migration Inhibitory Factor (MIF) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Macrophage Migration Inhibitory Factor (MIF) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-92 82
EDTA plasma(n=5) 80-92 81
heparin plasma(n=5) 97-104 101
sodium citrate plasma(n=5) 95-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Macrophage Migration Inhibitory Factor (MIF) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Macrophage Migration Inhibitory Factor (MIF) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Macrophage Migration Inhibitory Factor (MIF) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 85-99% 78-91% 92-102% 85-97%
EDTA plasma(n=5) 91-98% 81-96% 78-94% 86-95%
heparin plasma(n=5) 80-90% 87-95% 88-101% 91-98%
sodium citrate plasma(n=5) 79-104% 80-104% 99-105% 79-94%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:MIF) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Kidney International Targeted reduction of advanced glycation improves renal function in obesity PubMed: 21412218
The Journal of Immunology Macrophage Migration Inhibitory Factor Plays a Role in the Regulation of Microfold (M) Cell-Mediated Transport in the Gut Jimmunol: 5673
The Journal of Endocrinology Involvement of exercise-induced macrophage migration inhibitory factor in the prevention of fatty liver disease PubMed: PMC3757527
The Journal of Immunology Role of macrophage migration inhibitory factor in the regulatory T cell response of tumor-bearing mice PubMed: PMC3466372
Diabetologia Deletion of bone-marrow-derived receptor for AGEs (RAGE) improves renal function in an experimental mouse model of diabetes Pubmed:24957662
BioMed Research International The Potential Role of Polymethyl Methacrylate as a New Packaging Material for the Implantable Medical Device in the Bladder PubMed: 25705692
Diabetologia Combined NOX1/4 inhibition with GKT137831 in mice provides dose-dependent reno- and atheroprotection even in established micro- and macrovascular disease. pubmed:28160092
Life Sciences Protective effect of chlorogenic acid on the inflammatory damage of pancreas and lung in mice with l-arginine-induced pancreatitis pubmed:28919396
Scientific Reports Cytokine MIF Enhances Blood-Brain Barrier Permeability: Impact for Therapy in Ischemic Stroke Pubmed:29335619
Ocular Immunology and Inflammation Immune Response and Mechanisms of IFN-γ in Administration for Keratomycosis Pubmed: 30307777
International Journal of Immunopathology and Pharmacology Effect of voluntary running on expression of myokines in brains of rats with depression Pubmed: 30834799
Biomarkers in Heart Failure and Associated Diseases Biomarkers in Heart Failure and Associated Diseases
Science China-Life Sciences Autophagy mediates the secretion of macrophage migration inhibitory factor from cardiomyocytes upon serum-starvation Pubmed: 31209799
Folia Morphologica Beneficial effects of voluntary over forced exercise on skeletal muscle structure and myokines expression Pubmed: 31802473
Biomed Pharmacother Deletion of macrophage migration inhibitory factor ameliorates inflammation in mice model severe acute pancreatitis Pubmed: 32062385
Sci Rep Association between MIF gene promoter rs755622 and susceptibility to coronary artery disease and inflammatory cytokines in the Chinese Han population 33850223
Commun Biol Macrophage migration inhibitory factor is overproduced through EGR1 in TET2low resting monocytes Pubmed:35115654
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