Biotin Conjugated Human Nectin-2 Recombinant Rabbit Monoclonal Antibody [PSH12-52] - Detector

Sandwich ELISA analysis of human Nectin-2 matched pair antibodies

Capture: HA723460, Human Nectin-2 Rabbit mAb [PSH11-54]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723460) diluted in carbonate/bicarbonate buffer, at a concentration of 1 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 Nectin-2 protein (HA210905) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723461, Biotin, 0.05 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.

Sandwich ELISA analysis of human Nectin-2 matched pair antibodies

Capture: HA723463, Human Nectin-2 Rabbit mAb [PSH12-53]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723463) diluted in carbonate/bicarbonate buffer, at a concentration of 1 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 Nectin-2 protein (HA210905) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723461, Biotin, 0.05 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 Nectin-2 in MCF7 and MOLT-4 extract samples based on a 1,000 µg/ml extract load.

Capture: HA723460, Human Nectin-2 Rabbit mAb [PSH11-54]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

The concentrations of Nectin-2 were measured in duplicates, interpolated from the Nectin-2 standard curve and corrected for sample dilution. Undiluted samples are MOLT-4 extract 50% and MCF7 extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nectin-2 concentration was determined to be 3,548 pg/ml in MCF7 extract and undetectable in MOLT-4 extract.

Interpolated concentrations of native Nectin-2 in MCF7 and MOLT-4 extract samples based on a 1,000 µg/ml extract load.

Capture: HA723463, Human Nectin-2 Rabbit mAb [PSH12-53]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

The concentrations of Nectin-2 were measured in duplicates, interpolated from the Nectin-2 standard curve and corrected for sample dilution. Undiluted samples are MOLT-4 extract 50% and MCF7 extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nectin-2 concentration was determined to be 4,062 pg/ml in MCF7 extract and undetectable in MOLT-4 extract.

Interpolated concentrations of spiked Nectin-2 in human cell culture media samples.

Capture: HA723460, Human Nectin-2 Rabbit mAb [PSH11-54]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

The concentrations of Nectin-2 were measured in duplicates, interpolated from the Nectin-2 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 Nectin-2 in human cell culture media samples.

Capture: HA723463, Human Nectin-2 Rabbit mAb [PSH12-53]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

The concentrations of Nectin-2 were measured in duplicates, interpolated from the Nectin-2 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).