Serum, plasma and other biological fluids
13.717-10000pg/mL The standard curve concentrations used for the ELISA's were 10000pg/mL, 3333.33pg/mL, 1111.11pg/mL, 370.37pg/mL, 123.457pg/mL, 41.152pg/mL, 13.717pg/mL
Sensitivity The minimum detectable dose of this kit is typically less than 6.4pg/mL. Specificity This assay has high sensitivity and excellent specificity for detection of Macrophage Migration Inhibitory Factor (MIF).
No significant cross-reactivity or interference between Macrophage Migration Inhibitory Factor (MIF) and analogues was observed.
|Matrix||Recovery range (%)||Average(%)|
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
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. Test principle The microtiter plate provided in this kit has been pre-coated with an antigen. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated secondary antibody. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the Macrophage Migration Inhibitory Factor (MIF) level in the sample or standard. Assay procedure summary 1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37oC;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 100µL Substrate Solution. Incubate 10 minutes at 37oC;
8. Read RLU value immediately.