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Sandwich ELISA analysis of Mouse E-Cadherin matched pair antibodies
Capture: HA723911, Mouse/Rat E-Cadherin Rabbit mAb [PSH17-24]
Detector: HA723912, Mouse/Rat E-Cadherin Rabbit mAb [PSH17-25]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723911) 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 Mouse E-Cadherin protein starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA723912, 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.
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Interpolated concentrations of native E-Cadherin in Mouse, Rat serum
and NIH/3T3 supernatant.
Capture: HA723911, Mouse/Rat E-Cadherin Rabbit mAb [PSH17-24]
Detector: HA723912, Mouse/Rat E-Cadherin Rabbit mAb [PSH17-25]
Interpolated concentration of native E-Cadherin was measured in duplicate at different sample concentrations and interpolated from the E-Cadherin standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean E-Cadherin concentration was determined to be 960,793 pg/ml in Mouse serum 257,170 pg/ml in Rat serum and undetermined in NIH/3T3 supernatant.
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Interpolated concentrations of native E-Cadherin in mouse liver homogenate extract samples based on a 1000 µg/ml extract load.
Capture: HA723911, Mouse/Rat E-Cadherin Rabbit mAb [PSH17-24]
Detector: HA723912, Mouse/Rat E-Cadherin Rabbit mAb [PSH17-25]
Interpolated concentration of native E-Cadherin was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean E-Cadherin concentration was determined to be 14,881 pg/mL in mouse liver homogenate extract.
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Interpolated concentrations of spiked E-Cadherin in cell culture media samples.
Capture: HA723911, Mouse/Rat E-Cadherin Rabbit mAb [PSH17-24]
Detector: HA723912, Mouse/Rat E-Cadherin Rabbit mAb [PSH17-25]
The concentrations of E-Cadherin were measured in duplicates, interpolated from the E-Cadherin 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).
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