Human FGF-21 ELISA kit
FGF21 is a metabolic hormone mainly produced in liver. It regulates glucose and lipid metabolism through pleiotropic actions in multiple tissues, and protects against metabolic damages under various stresses.
In humans, high circulating FGF21 levels are found in obesity and multiple cardiometabolic disorders, including metabolic syndrome, type 2 diabetes, non-alcoholic fatty liver disease and coronary artery disease. Serum FGF21 is a potential biomarker for the early detection and risk prediction of these diseases.
PRINCIPLE OF THE ASSAY
This assay is a quantitative sandwich ELISA. The immunoplate is pre-coated with a rabbit polyclonal antibody specific for human FGF-21. Standards and samples are pipetted into the wells and any human FGF-21 present is bound by the immobilized antibody. After washing away any unbound substances, a biotin labelled polyclonal antibody specific for human FGF-21 is added to the wells. After wash step to remove any unbound reagents, streptavidin-HRP conjugate (STP-HRP) is added. After the last wash step, an HRP substrate solution is added and colour develops in proportion to the amount of human FGF-21 bound initially. The assay is stopped and the optical density of the wells determined using a microplate reader. Since the increases in absorbance are directly proportional to the amount of captured human FGF-21, the unknown sample concentration can be interpolated from a reference curve included in each assay.
A. Typical representation of standard curve
The following standard curve is provided for demonstration only. A standard curve should be generated for each set of sample assay.
Human FGF-21 (pg/ml)
The lowest level of human FGF-21 that can be detected by this assay is 15 pg/ml.
Cross Reactivity of recombinant proteins
Intra-assay Precision (Precision within an assay)
Two samples of known concentration were tested 12 times on one plate.
Inter-assay Precision (Precision between assays)
Two samples of known concentration were tested in 10 separate assays.
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human FGF-21 were serially diluted with the 1×Assay buffer to produce samples with values within the dynamic range of the assay.