Assay Principle

This kit employs double antibody sandwich ELISA technology: Capture Antibody is coated on the microplate to capture GCDH from samples and standards. After washing, biotin-labeled Detection Antibody is added and incubated, followed by washing to form a "Capture Antibody-Antigen-Detection Antibody" immune-complex. Subsequently, Streptavidin-Horseradish Peroxidase (SA-HRP) is added and incubated. After incubation and washing, TMB Substrate is added for color development. If the target analyte is present in the sample, a blue color develops. Stop Solution is then added to terminate the reaction. During the assay, unbound components are washed away. The Optical Density (OD) is measured at 450 nm using a microplate reader. The intensity of the color is proportional to the GCDH concentration in the sample, and the concentration is calculated by plotting a standard curve.

 

Typical Data

The following data and curve are for reference only. The experimenter must establish a standard curve based on their own experimental data.

Standard Conc. (pg/mL)	2000	1000	500	250	125	62.5	0
OD Value	2.704	1.533	0.794	0.423	0.221	0.134	0.037
Corrected OD Value	2.667	1.496	0.757	0.386	0.184	0.097	0

 

Recovery

Recovery was tested by spiking known concentrations of Human GCDH into different sample matrices. The recovery range and average recovery are shown below.

Sample Type	Range (%)	Average Recovery (%)
Cultured Cells (n=8)	92-103	97
Tissue Samples (n=8)	96-115	107

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