Biotin Conjugated Human DPP4 / CD26 Recombinant Rabbit Monoclonal Antibody [PSH19-27] - Detector

Sandwich ELISA analysis of Human DPP4 matched pair antibodies

Capture: HA724058, Human DPP4 / CD26 Rabbit mAb [PSH19-26]
Detector: HA724059, Human DPP4 / CD26 Rabbit mAb [PSH19-27]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724058) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 DPP4 / CD26 protein (HA211034) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA724059, 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.

Sandwich ELISA analysis of Human DPP4 matched pair antibodies

Capture: HA724060, Human DPP4 / CD26 Rabbit mAb [PSH19-28]
Detector: HA724059, Human DPP4 / CD26 Rabbit mAb [PSH19-27]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724060) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 DPP4 / CD26 protein (HA211034) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA724059, 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 DPP4 in HepG2,MCF-7 culture supernatant and Human urine.

Capture: HA724058, Human DPP4 / CD26 Rabbit mAb [PSH19-26]
Detector: HA724059, Human DPP4 / CD26 Rabbit mAb [PSH19-27]

Interpolated concentration of native DPP4 was measured in duplicate at different sample concentrations and interpolated from the DPP4 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean DPP4 concentration was determined to be 25,806 pg/ml in Human urine,4,573 pg/ml in HepG2 cell culture supernatant and undetectable in MCF-7 cell culture supernatant.

Interpolated concentrations of native DPP4 in HepG2,MCF-7 culture supernatant and Human urine.

Capture: HA724060, Human DPP4 / CD26 Rabbit mAb [PSH19-28]
Detector: HA724059, Human DPP4 / CD26 Rabbit mAb [PSH19-27]

Interpolated concentration of native DPP4 was measured in duplicate at different sample concentrations and interpolated from the DPP4 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean DPP4 concentration was determined to be 25,703 pg/ml in Human urine,5,564 pg/ml in HepG2 cell culture supernatant and undetectable in MCF-7 cell culture supernatant.

Interpolated concentrations of spiked DPP4 in cell culture media samples.

Capture: HA724058, Human DPP4 / CD26 Rabbit mAb [PSH19-26]
Detector: HA724059, Human DPP4 / CD26 Rabbit mAb [PSH19-27]

The concentrations of DPP4 were measured in duplicates, interpolated from the DPP4 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 DPP4 in cell culture media samples.

Capture: HA724060, Human DPP4 / CD26 Rabbit mAb [PSH19-28]
Detector: HA724059, Human DPP4 / CD26 Rabbit mAb [PSH19-27]

The concentrations of DPP4 were measured in duplicates, interpolated from the DPP4 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).