Details
Assay Principle
This kit employs double antibody sandwich ELISA technology: Capture Antibody is coated on the microplate to capture VEGF-A 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 VEGF-A 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 |
31.25 |
0 |
|
2.703 |
1.534 |
0.793 |
0.423 |
0.225 |
0.134 |
0.087 |
0.035 |
|
|
Corrected OD Value |
2.668 |
1.499 |
0.758 |
0.388 |
0.19 |
0.099 |
0.052 |
0 |
Sensitivity
The minimum detectable concentration (sensitivity) of Human VEGF-A, determined by testing samples, is 15.625pg/mL.
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