Biotin Conjugated Human EGFR Recombinant Rabbit Monoclonal Antibody [PSH19-10] - Detector

Sandwich ELISA analysis of Human EGFR matched pair antibodies

Capture: HA724043, Human EGFR Rabbit mAb [PSH19-09]
Detector: HA724044, Human EGFR Rabbit mAb [PSH19-10]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724043) 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 EGFR protein (HA211104) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA724044, 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 EGFR in A431 and Jurkat cell
culture supernatant.

Capture: HA724043, Human EGFR Rabbit mAb [PSH19-09]
Detector: HA724044, Human EGFR Rabbit mAb [PSH19-10]

Interpolated concentration of native EGFR was measured in duplicate at different sample concentrations and interpolated from the EGFR standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean EGFR concentration was determined to be 84,020 pg/ml in A431 cell culture supernatant and undetectable in Jurkat cell culture supernatant.

Interpolated concentrations of spiked EGFR in cell culture media samples.

Capture: HA724043, Human EGFR Rabbit mAb [PSH19-09]
Detector: HA724044, Human EGFR Rabbit mAb [PSH19-10]

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