Biotin Conjugated Human Livin Recombinant Rabbit Monoclonal Antibody [PSH12-95] - Detector

Sandwich ELISA analysis of Human Livin matched pair antibodies

Capture: HA723498, Human Livin Rabbit mAb [PSH12-94]
Detector: HA723499, Human Livin Rabbit mAb [PSH12-95]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723498) 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 Angiogenin protein (HA211325) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723499, 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 Livin matched pair antibodies

Capture: HA723501, Human Livin Rabbit mAb [PSH12-96]
Detector: HA723499, Human Livin Rabbit mAb [PSH12-95]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723501) 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 Angiogenin protein (HA211325) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723499, 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 Livin in SK-MEL-28 and HeLa extract samples based on a 1000 µg/ml extract load.

Capture: HA723498, Human Livin Rabbit mAb [PSH12-94]
Detector: HA723499, Human Livin Rabbit mAb [PSH12-95]

Interpolated concentration of native Livin was measured in duplicate at different sample concentrations and interpolated from the Livin standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Livin concentration was determined to be 134,381 pg/mL in SK-MEL-28 cell extract. There was no detectable signal in HeLa cell extract.

Interpolated concentrations of native Livin in SK-MEL-28 and HeLa extract samples based on a 1000 µg/ml extract load.

Capture: HA723501, Human Livin Rabbit mAb [PSH12-96]
Detector: HA723499, Human Livin Rabbit mAb [PSH12-95]

Interpolated concentration of native Livin was measured in duplicate at different sample concentrations and interpolated from the Livin standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Livin concentration was determined to be 53,202 pg/mL in SK-MEL-28 cell extract. There was no detectable signal in HeLa cell extract.

Interpolated concentrations of spiked Livin in cell culture media samples.

Capture: HA723498, Human Livin Rabbit mAb [PSH12-94]
Detector: HA723499, Human Livin Rabbit mAb [PSH12-95]

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

Capture: HA723501, Human Livin Rabbit mAb [PSH12-96]
Detector: HA723499, Human Livin Rabbit mAb [PSH12-95]

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