Biotin Conjugated Human VEGFA Recombinant Rabbit Monoclonal Antibody [PSH12-93] - Detector

Sandwich ELISA analysis of Human VEGFA matched pair antibodies

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723496, Human VEGFA Rabbit mAb [PSH12-93]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723493) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 VEGFA protein (HA210929) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723496, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 VEGFA in A431 and HepG2 cell culture supernatant.

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723496, Human VEGFA Rabbit mAb [PSH12-93]

The concentrations of VEGFA measured in duplicate and interpolated from the VEGFA standard curve and corrected for sample dilution. Undiluted samples are as follows: A431 cell culture supernatant 12.5% and HepG2 cell culture supernatant 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGFA concentration was determined to be 6,139 pg/ml in A431 cell culture supernatant and 3,773 pg/ml in HepG2 cell culture supernatant.

Interpolated concentrations of spiked VEGF in human cell culture media samples.

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723496, Human VEGFA Rabbit mAb [PSH12-93]

The concentrations of VEGFA were measured in duplicates, interpolated from the VEGFA standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).