Biotin Conjugated Human PD-L2 Recombinant Rabbit Monoclonal Antibody [PSH09-82] - Detector

Sandwich ELISA analysis of Human PD-L2 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723145) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 Human PD-L2 (HA210599) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA723148B, 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.

Sandwich ELISA analysis of Human PD-L2 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723146) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 Human PD-L2 (HA210599) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA723148B, 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 PD-L2 in human urine, human serum samples and Jurkat cell culture supernatant.

The concentrations of PD-L2 were measured in duplicate, interpolated from the PD-L2 standard curve and corrected for sample dilution. Undiluted samples are human urine 100%, human serum 40%, and Jurkat cell culture supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PD-L2 concentration was determined to be 4,248 pg/ml in human urine, 10,104 pg/ml in human serum and undetectable in Jurkat cell culture supernatant.

Interpolated concentrations of native PD-L2 in human urine, human serum samples and Jurkat cell culture supernatant.

The concentrations of PD-L2 were interpolated from the PD-L2 standard curve and corrected for sample dilution. Undiluted samples are human urine 100%, human serum 40%, and Jurkat cell culture supernatant 100%. The mean PD-L2 concentration was determined to be 4,983 pg/ml in human urine, 14,438 pg/ml in human serum and undetectable in Jurkat cell culture supernatant.

Interpolated concentrations of spiked PD-L2 in human cell culture media samples.

The concentrations of PD-L2 were measured in duplicates, interpolated from the PD-L2 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).

Interpolated concentrations of spiked PD-L2 in human cell culture media samples.

The concentrations of PD-L2 were measured in duplicates, interpolated from the PD-L2 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).