The human DKK1 ELISA kit is a 3.5 hour, 96-well sandwich ELISA for the quantitative determination of DKK-1 in human serum. The assay employs human serum-based standards to ensure the measurement of biologically reliable data.
DKK1 ELISA Assay Principle
The DKK-1 ELISA kit is a sandwich enzyme immunoassay for the quantitative determination of DKK1 in human serum samples.
The figure below explains the principle of the DKK1 ELISA assay:
In a first step, assay buffer, standard/control/sample and detection antibody (biotinylated polyclonal rabbit anti-human DKK-1) are pipetted into the wells of the microtiter strips, which are pre-coated with monoclonal mouse anti-human DKK-1 antibody. DKK-1 present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody. In the washing step all non-specific unbound material is removed. In a next step, the conjugate (streptavidin-HRP) is pipetted into the wells and reacts with the detection antibody. After another washing step, the substrate (TMB, tetramethylbenzidine) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of DKK-1 present in the sample. This color change is detectable with a standard microtiter plate reader. A dose response curve of the absorbance (optical density, OD at 450 nm) versus standard concentration is generated using the values obtained from the standard. The concentration of DKK-1 in the sample is determined directly from the dose response curve.
DKK1 ELISA Typical Standard Curve
The figure below shows a typical standard curve for the Dickkopf-1 ELISA.The immunoassay is calibrated againstrecombinant human DKK1.
DKK1 ELISA Kit Components
Storage instructions:All reagents of the DKK-1 ELISA kit are stable at 4°C (2-8°C) until the expiry date stated on the label of each reagent.
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