Human Antithrombin III / ATIII Recombinant Rabbit Monoclonal Antibody [PSH14-25] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human Antithrombin III matched pair antibodies

Capture: HA723615, Human Antithrombin III Rabbit mAb [PSH14-25]
Detector: HA723616, Human Antithrombin III Rabbit mAb [PSH14-26]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723615) 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 Antithrombin III protein (HA210985) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA723616, 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 Antithrombin III in HepG2
culture supernatant and Human urine.

Capture: HA723615, Human Antithrombin III Rabbit mAb [PSH14-25]
Detector: HA723616, Human Antithrombin III Rabbit mAb [PSH14-26]

Interpolated concentration of native Antithrombin III was measured in duplicate at different sample concentrations and interpolated from the Antithrombin III standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Antithrombin III concentration was determined to be 25046 pg/ml in HepG2 cell culture supernatant,317 pg/ml in Human urine.

Interpolated concentrations of spiked Antithrombin III in cell culture media samples.

Capture: HA723615, Human Antithrombin III Rabbit mAb [PSH14-25]
Detector: HA723616, Human Antithrombin III Rabbit mAb [PSH14-26]

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