This assay has high sensitivity and excellent specificity for detection of Fibroblast Growth Factor 21 (FGF21).No significant cross-reactivity or interference between Fibroblast Growth Factor 21 (FGF21) and analogues was observed.

Matrices listed below were spiked with certain level of recombinant Fibroblast Growth Factor 21 (FGF21) and the recovery rates were calculated by comparing the measured value to the expected amount of Fibroblast Growth Factor 21 (FGF21) in samples.

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibroblast Growth Factor 21 (FGF21) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Fibroblast Growth Factor 21 (FGF21) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10%>Inter-Assay: CV<12%>

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Fibroblast Growth Factor 21 (FGF21) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

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.

The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to Fibroblast Growth Factor 21 (FGF21). A competitive inhibition reaction is launched between biotin labeled Fibroblast Growth Factor 21 (FGF21) and unlabeled Fibroblast Growth Factor 21 (FGF21) (Standards or samples) with the pre-coated antibody specific to Fibroblast Growth Factor 21 (FGF21). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Fibroblast Growth Factor 21 (FGF21) in the sample. 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 reverse proportional to the Fibroblast Growth Factor 21 (FGF21) level in the sample or standard.

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