Human Prostate Specific Antigen Recombinant Rabbit Monoclonal Antibody [PSH20-05] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human Prostate Specific Antigen matched pair antibodies

Capture: HA725257, Human Prostate Specific Antigen Rabbit mAb [PSH20-04]
Detector: HA725258, Human Prostate Specific Antigen Rabbit mAb [PSH20-05]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725257) 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 Prostate Specific Antigen protein (HA211208) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA725258, 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 Prostate Specific Antigen in Lncap and DU 145 cell culture supernatant.

Capture: HA725257, Human Prostate Specific Antigen Rabbit mAb [PSH20-04]
Detector: HA725258, Human Prostate Specific Antigen Rabbit mAb [PSH20-05]

The concentrations of Prostate Specific Antigen were measured in triplicates, interpolated from the Prostate Specific Antigen standard curve and corrected for sample dilution. Undiluted samples are Lncap cell culture supernatant 2% and DU 145 cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean Prostate Specific Antigen concentration was determined to be 30,935 pg/ml in Lncap and undetectable in DU 145 cell culture supernatant.