Biotin Conjugated Human PD1 Recombinant Rabbit Monoclonal Antibody [PSH13-38] - Detector

Sandwich ELISA analysis of Human PD1 matched pair antibodies

Capture: HA723521, Human PD1 Rabbit mAb [SJ01-91]
Detector: HA723522, Human PD1 Rabbit mAb [PSH13-38]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723521) 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 PD1 protein (HA210922) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA723522, 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 PD1 in MOLT-4 and HeLa extract samples based on a 1,000 µg/ml extract load.

Capture: HA723521, Human PD1 Rabbit mAb [SJ01-91]
Detector: HA723522, Human PD1 Rabbit mAb [PSH13-38]

The concentrations of PD1 were measured in triplicates, interpolated from the PD1 standard curve and corrected for sample dilution. Undiluted samples are MOLT-4 extract 100% and HeLa extract 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean PD1 concentration was determined to be 148.6 pg/ml in MOLT-4 extract and undetectable in HeLa extract.

Interpolated concentrations of native PD1 in human urine samples.

Capture: HA723521, Human PD1 Rabbit mAb [SJ01-91]
Detector: HA723522, Human PD1 Rabbit mAb [PSH13-38]

The concentrations of PD1 were measured in duplicates, interpolated from the PD1 standard curve and corrected for sample dilution. Undiluted samples are human urine 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PD1 concentration was determined to be 243.4 pg/ml in human urine.

Interpolated concentrations of spiked PD1 in human cell culture media samples.

Capture: HA723521, Human PD1 Rabbit mAb [SJ01-91]
Detector: HA723522, Human PD1 Rabbit mAb [PSH13-38]

The concentrations of PD1 were measured in duplicates, interpolated from the PD1 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).