Human Caspase-9 Recombinant Rabbit Monoclonal Antibody [PSH17-20] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human Caspase-9 matched pair antibodies

Capture: HA723906, Human Caspase-9 Rabbit mAb [PSH17-19]
Detector: HA723907, Human Caspase-9 Rabbit mAb [PSH17-20]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723906) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 Caspase-9 protein (HA211015) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723907, 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 caspase-9 in THP-1, Hela and Jurkat cell extract samples based on a 1000 µg/ml extract load.

Capture: HA723906, Human Caspase-9 Rabbit mAb [PSH17-19]
Detector: HA723907, Human Caspase-9 Rabbit mAb [PSH17-20]

Interpolated concentration of native caspase-9 was measured in duplicate at different sample concentrations and interpolated from the caspase-9 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean caspase-9 concentration was determined to be 6,717 pg/mL in THP-1, 6,267 pg/mL in Hela and 3,257 pg/mL in Jurkat cell extract.

Interpolated concentrations of spiked caspase-9 in cell culture media samples.

Capture: HA723906, Human Caspase-9 Rabbit mAb [PSH17-19]
Detector: HA723907, Human Caspase-9 Rabbit mAb [PSH17-20]

The concentrations of caspase-9 were measured in duplicates, interpolated from the caspase-9 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).