Human alpha Fetoprotein Recombinant Rabbit Monoclonal Antibody [PSH14-53] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human alpha Fetoprotein matched pair antibodies

Capture: HA723644, Human alpha Fetoprotein Rabbit mAb [PSH14-53]
Detector: HA723645, Human alpha Fetoprotein Rabbit mAb [PSH14-54]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723644) 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 alpha Fetoprotein protein (HA210967) starting from 10000 pg/ml to 0 pg/ml and detect antibody (HA723645, 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 alpha Fetoprotein matched pair antibodies

Capture: HA723644, Human alpha Fetoprotein Rabbit mAb [PSH14-53]
Detector: HA723645, Human alpha Fetoprotein Rabbit mAb [PSH14-55]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723644) 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 alpha Fetoprotein protein (HA210967) starting from 10000 pg/ml to 0 pg/ml and detect antibody (HA723647, 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 Alpha-fetoprotein in human samples.

Capture: HA723644, Human alpha Fetoprotein Rabbit mAb [PSH14-53]
Detector: HA723645, Human alpha Fetoprotein Rabbit mAb [PSH14-54]

Interpolated concentration of native Alpha-fetoprotein was measured in duplicate at different sample concentrations and interpolated from the Alpha-fetoprotein standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Alpha-fetoprotein concentration was determined to be 1.49 ug/mL in Hep G2 cell supernatant. There was no detectable signal in THP-1 cell supernatant.

Interpolated concentrations of native Alpha-fetoprotein in human samples.

Capture: HA723644, Human alpha Fetoprotein Rabbit mAb [PSH14-53]
Detector: HA723645, Human alpha Fetoprotein Rabbit mAb [PSH14-55]

Interpolated concentration of native Alpha-fetoprotein was measured in duplicate at different sample concentrations and interpolated from the Alpha-fetoprotein standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Alpha-fetoprotein concentration was determined to be 1.8 ug/mL in Hep G2 cell supernatant. There was no detectable signal in THP-1 cell supernatant.

Interpolated concentrations of spiked Alpha-fetoprotein in cell culture media samples.

Capture: HA723644, Human alpha Fetoprotein Rabbit mAb [PSH14-53]
Detector: HA723645, Human alpha Fetoprotein Rabbit mAb [PSH14-54]

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

Capture: HA723644, Human alpha Fetoprotein Rabbit mAb [PSH14-53]
Detector: HA723645, Human alpha Fetoprotein Rabbit mAb [PSH14-55]

The concentrations of Alpha-fetoprotein were measured in duplicates, interpolated from the Alpha-fetoprotein 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).