Biotin Conjugated Human VEGFD Recombinant Rabbit Monoclonal Antibody [PSH25-12] - Detector

Sandwich ELISA analysis of Human VEGFD matched pair antibodies

Capture: HA724505, Human VEGFD Rabbit mAb [PSH25-11]
Detector: HA724506, Human VEGFD Rabbit mAb [PSH25-12]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724505) diluted in carbonate/bicarbonate buffer, at a concentration of 2 μg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human VEGFD protein (HA211253) starting from 3,000 pg/mL to 0 pg/mL and detect antibody (HA724506, Biotin, 0.2 μg/mL) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Interpolated concentrations of native VEGFD in human serum.

Capture: HA724505, Human VEGFD Rabbit mAb [PSH25-11]
Detector: HA724506, Human VEGFD Rabbit mAb [PSH25-12]

Interpolated concentration of native VEGFD was measured in duplicate at different sample concentrations and interpolated from the VEGFD standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean VEGFD concentration was determined to be 412.5 pg/mL in human serum.

Interpolated concentrations of spiked VEGFD in cell culture media samples.

Capture: HA724505, Human VEGFD Rabbit mAb [PSH25-11]
Detector: HA724506, Human VEGFD Rabbit mAb [PSH25-12]

The concentrations of VEGFD were measured in duplicates, interpolated from the VEGFD standard curves and corrected for sample dilution. diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).