P-Selectin / CD62P Recombinant Rabbit Monoclonal Antibody [PSH16-70] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human P-Selectin matched pair antibodies

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723872) 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 P-Selectin / CD62P protein (HA210931) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA723873, 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 mouse P-Selectin matched pair antibodies

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723872) 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 Mouse P-Selectin / CD62P protein (HA211140) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA723873, 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 P-Selectin in human serum samples and HeLa cell culture supernatant.

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

The concentrations of P-Selectin were measured in triplicates, interpolated from the P-Selectin standard curve and corrected for sample dilution. Undiluted samples are human serum 5% and HeLa cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean P-Selectin concentration was determined to be 95,663 pg/ml in human serum and undetectable in HeLa cell culture supernatant.

Interpolated concentrations of native P-Selectin in mouse serum samples.

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

The concentrations of P-Selectin were measured in duplicates, interpolated from the P-Selectin standard curve and corrected for sample dilution. Undiluted samples are mouse serum 1.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean P-Selectin concentration was determined to be 849,328 pg/ml in mouse serum.

Interpolated concentrations of native P-Selectin in rat serum samples.

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

The concentrations of P-Selectin were measured in duplicates, interpolated from the P-Selectin standard curve and corrected for sample dilution. Undiluted samples are rat serum 1.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean P-Selectin concentration was determined to be 118,074 pg/ml in rat serum.

Interpolated concentrations of spiked P-Selectin in human cell culture media samples.

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

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

Interpolated concentrations of spiked P-Selectin in mouse cell culture media samples.

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

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