Human Fetuin A Recombinant Rabbit Monoclonal Antibody [PSH19-12] - BSA and Azide free (Capture/Detector)

Sandwich ELISA analysis of Human Fetuin A matched pair antibodies

Capture: HA724046, Human Fetuin A Rabbit mAb [PSH19-12]
Detector: HA724047, Human Fetuin A Rabbit mAb [PSH19-13]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724046) 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 Fetuin A protein (HA211146) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA724047, 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 Fetuin A in Human serum.

Capture: HA724046, Human Fetuin A Rabbit mAb [PSH19-12]
Detector: HA724047, Human Fetuin A Rabbit mAb [PSH19-13]

Interpolated concentration of native Fetuin A was measured in duplicate at different sample concentrations and interpolated from the Fetuin A standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Fetuin A concentration was determined to be 1,006 ug/ml in Human serum.

Interpolated concentrations of native Fetuin A in HepG2 and A549 cell
culture supernatant.

Capture: HA724046, Human Fetuin A Rabbit mAb [PSH19-12]
Detector: HA724047, Human Fetuin A Rabbit mAb [PSH19-13]

Interpolated concentration of native Fetuin A was measured in duplicate at different sample concentrations and interpolated from the Fetuin A standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Fetuin A concentration was determined to be 1.2 ug/ml in HepG2 cell culture supernatant and undetectable in A549 cell culture supernatant.

Interpolated concentrations of spiked Fetuin A in cell culture media samples.

Capture: HA724046, Human Fetuin A Rabbit mAb [PSH19-12]
Detector: HA724047, Human Fetuin A Rabbit mAb [PSH19-13]

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

Sandwich ELISA analysis of Human Fetuin A matched pair antibodies

Capture: HA724048, Human Fetuin A Rabbit mAb [PSH19-14]
Detector: HA724046, Human Fetuin A Rabbit mAb [PSH19-12]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724048) 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 Fetuin A protein (HA211146) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA724046, 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 Fetuin A in Human serum.

Capture: HA724048, Human Fetuin A Rabbit mAb [PSH19-14]
Detector: HA724046, Human Fetuin A Rabbit mAb [PSH19-12]

Interpolated concentration of native Fetuin A was measured in duplicate at different sample concentrations and interpolated from the Fetuin A standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2).The mean Fetuin A concentration was determined to be 642 ug/ml in Human serum.

Interpolated concentrations of native Fetuin A in HepG2 and A549 cell
culture supernatant.

Capture: HA724048, Human Fetuin A Rabbit mAb [PSH19-14]
Detector: HA724046, Human Fetuin A Rabbit mAb [PSH19-12]

Interpolated concentration of native Fetuin A was measured in duplicate at different sample concentrations and interpolated from the Fetuin A standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Fetuin A concentration was determined to be 1.1 ug/ml in HepG2 cell culture supernatant and undetectable in A549 cell culture supernatant.

Interpolated concentrations of spiked Fetuin A in cell culture media samples.

Capture: HA724048, Human Fetuin A Rabbit mAb [PSH19-14]
Detector: HA724046, Human Fetuin A Rabbit mAb [PSH19-12]

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