Biotin Conjugated Human ACE2 Recombinant Rabbit Monoclonal Antibody [PSH17-18] - Detector

Sandwich ELISA analysis of Human ACE2 matched pair antibodies

Capture: HA723904, Human ACE2 Rabbit mAb [PSH17-17]
Detector: HA723905, Human ACE2 Rabbit mAb [PSH17-18]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723904) 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 ACE2 protein (HAg2008) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA723905, 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 ACE2 in human serum samples and HeLa cell culture supernatant.

Capture: HA723904, Human ACE2 Rabbit mAb [PSH17-17]
Detector: HA723905, Human ACE2 Rabbit mAb [PSH17-18]

The concentrations of ACE2 were measured in duplicates, interpolated from the ACE2 standard curve and corrected for sample dilution. Undiluted samples are human serum 50% and HeLa cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ACE2 concentration was determined to be 1,633 pg/ml in human serum and undetectable in HeLa cell culture supernatant.

Interpolated concentrations of spiked ACE2 in human cell culture media samples.

Capture: HA723904, Human ACE2 Rabbit mAb [PSH17-17]
Detector: HA723905, Human ACE2 Rabbit mAb [PSH17-18]

The concentrations of ACE2 were measured in duplicates, interpolated from the ACE2 standard curves and corrected for sample dilution. Diluted samples are as follows: 25% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).