ELISA Kit for Macrophage Migration Inhibitory Factor (MIF) Rattus norvegicus (Rat) Sandwich ELISA

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

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  • ELISA Kit for Macrophage Migration Inhibitory Factor (MIF) Packages (Simulation)
  • ELISA Kit for Macrophage Migration Inhibitory Factor (MIF) Packages (Simulation)
  • ELISA Kit for Macrophage Migration Inhibitory Factor (MIF) Results demonstration
  • SEA698Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 82-99 87
EDTA plasma(n=5) 84-95 89
heparin plasma(n=5) 96-103 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Macrophage Migration Inhibitory Factor (MIF) 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) 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) 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) 82-96% 79-91% 85-101% 85-96%
EDTA plasma(n=5) 93-102% 87-101% 93-105% 80-97%
heparin plasma(n=5) 87-101% 95-102% 80-97% 94-102%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Macrophage Migration Inhibitory Factor (MIF)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Macrophage Migration Inhibitory Factor (MIF). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Macrophage Migration Inhibitory Factor (MIF). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Macrophage Migration Inhibitory Factor (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 Macrophage Migration Inhibitory Factor (MIF) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Targeted reduction of advanced glycation improves renal function in obesityPubMed: 21412218
  • Macrophage Migration Inhibitory Factor Plays a Role in the Regulation of Microfold (M) Cell-Mediated Transport in the GutJimmunol: 5673
  • Involvement of exercise-induced macrophage migration inhibitory factor in the prevention of fatty liver diseasePubMed: PMC3757527
  • Role of macrophage migration inhibitory factor in the regulatory T cell response of tumor-bearing micePubMed: PMC3466372
  • Deletion of bone-marrow-derived receptor for AGEs (RAGE) improves renal function in an experimental mouse model of diabetesPubmed:24957662
  • The Potential Role of Polymethyl Methacrylate as a New Packaging Material for the Implantable Medical Device in the BladderPubMed: 25705692
  • Combined NOX1/4 inhibition with GKT137831 in mice provides dose-dependent reno- and atheroprotection even in established micro- and macrovascular disease.pubmed:28160092
  • Protective effect of chlorogenic acid on the inflammatory damage of pancreas and lung in mice with l-arginine-induced pancreatitis pubmed:28919396
  • Cytokine MIF Enhances Blood-Brain Barrier Permeability: Impact for Therapy in Ischemic StrokePubmed:29335619
  • Immune Response and Mechanisms of IFN-γ in Administration for KeratomycosisPubmed: 30307777
  • Effect of voluntary running on expression of myokines in brains of rats with depressionPubmed: 30834799
  • Biomarkers in Heart Failure and Associated Diseases
  • Autophagy mediates the secretion of macrophage migration inhibitory factor from cardiomyocytes upon serum-starvationPubmed: 31209799
  • Beneficial effects of voluntary over forced exercise on skeletal muscle structure and myokines expressionPubmed: 31802473
  • Deletion of macrophage migration inhibitory factor ameliorates inflammation in mice model severe acute pancreatitisPubmed: 32062385
  • Association between MIF gene promoter rs755622 and susceptibility to coronary artery disease and inflammatory cytokines in the Chinese Han population33850223
  • Macrophage migration inhibitory factor is overproduced through EGR1 in TET2low resting monocytesPubmed:35115654

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