Human CD62E Recombinant Rabbit Monoclonal Antibody [PSH14-42] - BSA and Azide free (Capture/Detector)

Sandwich ELISA analysis of Human CD62E matched pair antibodies

Capture: HA723635, Human CD62E Rabbit mAb [PSH14-41]
Detector: HA723636, Human CD62E Rabbit mAb [PSH14-42]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723635) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 CD62E protein (HA210996) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA723636, 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 Human CD62E matched pair antibodies

Capture: HA723636, Human CD62E Rabbit mAb [PSH14-42]
Detector: HA723638, Human CD62E Rabbit mAb [PSH14-43]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723636) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 CD62E protein (HA210996) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA723638, 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 CD62E in human samples.

Capture: HA723635, Human CD62E Rabbit mAb [PSH14-41]
Detector: HA723636, Human CD62E Rabbit mAb [PSH14-42]

Interpolated concentration of native CD62E was measured in duplicate at different sample concentrations and interpolated from the CD62E standard curves. Undiluted samples were 5% human serum and 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD62E concentration was determined to be 43，079 pg/mL in human serum. There was no detectable signal in MCF-7 cell supernatant.

Interpolated concentrations of native CD62E in human samples.

Capture: HA723636, Human CD62E Rabbit mAb [PSH14-42]
Detector: HA723638, Human CD62E Rabbit mAb [PSH14-43]

Interpolated concentration of native CD62E was measured in duplicate at different sample concentrations and interpolated from the CD62E standard curves. Undiluted samples were 5% human serum and 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD62E concentration was determined to be 23,360 pg/mL in human serum. There was no detectable signal in MCF-7 cell supernatant.

Interpolated concentrations of spiked CD62E in cell culture media samples.

Capture: HA723635, Human CD62E Rabbit mAb [PSH14-41]
Detector: HA723636, Human CD62E Rabbit mAb [PSH14-42]

The concentrations of CD62E were measured in duplicates, interpolated from the CD62E 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 CD62E in cell culture media samples.

Capture: HA723636, Human CD62E Rabbit mAb [PSH14-42]
Detector: HA723638, Human CD62E Rabbit mAb [PSH14-43]

The concentrations of CD62E were measured in duplicates, interpolated from the CD62E 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).