Human Fas / CD95 Recombinant Rabbit Monoclonal Antibody [PSH16-37] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human Fas / CD95 matched pair antibodies

Capture: HA723837, Human Fas / CD95 Rabbit mAb [PSH16-39]
Detector: HA723835, Human Fas / CD95 Rabbit mAb [PSH16-37]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723837) diluted in carbonate/bicarbonate buffer, at a concentration of 2 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 Fas / CD95 protein (HA211064) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA723835, 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 FAS in Raji,Ramos and K-562 Cell extract samples based on a 1000 µg/ml extract load.

Capture: HA723837, Human Fas / CD95 Rabbit mAb [PSH16-39]
Detector: HA723835, Human Fas / CD95 Rabbit mAb [PSH16-37]

Interpolated concentration of native FAS was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean FAS concentration was determined to be 15,453 pg/ml in Raji Cell extract 529 pg/ml in Ramos Cell extract and undetectable in K-562 Cell extract.

Interpolated concentrations of spiked FAS in cell culture media samples.

Capture: HA723837, Human Fas / CD95 Rabbit mAb [PSH16-39]
Detector: HA723835, Human Fas / CD95 Rabbit mAb [PSH16-37]

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