Biotin Conjugated Human Poliovirus Receptor/PVR Recombinant Rabbit Monoclonal Antibody [PSH12-55] - Detector

Sandwich ELISA analysis of Human Poliovirus Receptor/PVR matched pair antibodies

Capture: HA723464, Human Poliovirus Receptor/PVR Rabbit mAb [PSH12-54]
Detector: HA723465, Human Poliovirus Receptor/PVRn Rabbit mAb [PSH12-55]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723464) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Poliovirus Receptor/PVRn protein (HA210957) starting from 8,000 pg/ml to 0 pg/ml and detect antibody (HA723465, 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 PVR in human samples.

Capture: HA723464, Human Poliovirus Receptor/PVR Rabbit mAb [PSH12-54]
Detector: HA723465, Human Poliovirus Receptor/PVRn Rabbit mAb [PSH12-55]

Interpolated concentration of native PVR was measured in duplicate at different sample concentrations. Undiluted samples were 1% human serum. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean PVR concentration was determined to be 133,172 pg/mL in human serum.

Interpolated concentrations of native PVR in U-87 MG and Raji extract samples based on a 1000 µg/ml extract load.

Capture: HA723464, Human Poliovirus Receptor/PVR Rabbit mAb [PSH12-54]
Detector: HA723465, Human Poliovirus Receptor/PVRn Rabbit mAb [PSH12-55]

Interpolated concentration of native PVR was measured in duplicate at different sample concentrations and interpolated from the PVR standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean PVR concentration was determined to be 7,903 pg/mL in U-87 MG cell extract. There was no detectable signal in Raji cell extract.

Interpolated concentrations of spiked PVR in cell culture media samples.

Capture: HA723464, Human Poliovirus Receptor/PVR Rabbit mAb [PSH12-54]
Detector: HA723465, Human Poliovirus Receptor/PVRn Rabbit mAb [PSH12-55]

The concentrations of PVR were measured in duplicates, interpolated from the PVR 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).