Human E-Cadherin Recombinant Rabbit Monoclonal Antibody [PSH09-02] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human E-Cadherin matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723055) 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 E-Cadherin protein (HA210886) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA723056, Biotin, 0.2 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 E-Cadherin in Jurkat and MCF-7 cell culture supernatant.

The concentrations of E-Cadherin were measured in duplicates, interpolated from the E-Cadherin standard curve and corrected for sample dilution. Undiluted samples are MCF-7 cell culture supernatant 50% and Jurkat cell culture supernatnat 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean E-Cadherin concentration was undetectable in Jurkat cell culture supernatant and 13.7 ng/ml in MCF-7 cell culture supernatant.

Interpolated concentrations of native E-Cadherin in human urine and human serum samples.

The concentrations of E-Cadherin were measured in duplicates, interpolated from the E-Cadherin standard curve and corrected for sample dilution. Undiluted samples are human urine 13% and human serum 10%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean E-Cadherin concentration was determined to be 33.1 ng/ml in human urine and 137.0 ng/ml in human serum.

Interpolated concentrations of spiked E-Cadherin in human cell culture media samples.

The concentrations of E-Cadherin were measured in duplicates, interpolated from the E-Cadherin 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).