Human CA125/MUC16 Recombinant Rabbit Monoclonal Antibody [PSH23-20] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human CA125/MUC16 matched pair antibodies

Capture: HA724354, Human CA125/MUC16 Rabbit mAb [PSH23-19]
Detector: HA724355, Human CA125/MUC16 Rabbit mAb [PSH23-20]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724354) 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 Human CA125/MUC16 protein (HA211195) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA724355, 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 CA125/MUC16 in Hela cell culture supernatant.

Capture: HA724354, Human CA125/MUC16 Rabbit mAb [PSH23-19]
Detector: HA724355, Human CA125/MUC16 Rabbit mAb [PSH23-20]

Interpolated concentration of native CA125/MUC16 was measured in duplicate at different sample concentrations and interpolated from the CA125/MUC16 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CA125/MUC16 concentration was determined to be 3,098 pg/ml in Hela cell culture supernatant.

Interpolated concentrations of native CA125/MUC16 in human serum and OVCAR-3 cell culture supernatant.

Capture: HA724354, Human CA125/MUC16 Rabbit mAb [PSH23-19]
Detector: HA724355, Human CA125/MUC16 Rabbit mAb [PSH23-20]

Interpolated concentration of native CA125/MUC16 was measured in duplicate at different sample concentrations and interpolated from the CA125/MUC16 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CA125/MUC16 concentration was determined to be 631 pg/ml in OVCAR-3 cell culture supernatant and 231 pg/ml in Human serum.

Interpolated concentrations of native CA125/MUC16 in OVCAR-3 and Hela Cell extract samples based on a 1000 µg/ml extract load.

Capture: HA724354, Human CA125/MUC16 Rabbit mAb [PSH23-19]
Detector: HA724355, Human CA125/MUC16 Rabbit mAb [PSH23-20]

Interpolated concentration of native CA125/MUC16 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CA125/MUC16 concentration was determined to be 68,073 pg/ml in OVCAR-3 Cell extract and 98,672 pg/ml in OVCAR-3 Cell extract.

Interpolated concentrations of spiked CA125/MUC16 in cell culture media samples.

Capture: HA724354, Human CA125/MUC16 Rabbit mAb [PSH23-19]
Detector: HA724355, Human CA125/MUC16 Rabbit mAb [PSH23-20]

The concentrations of CA125/MUC16 were measured in duplicates, interpolated from the CA125/MUC16 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).