Biotin Conjugated Human Thrombopoietin Recombinant Rabbit Monoclonal Antibody [PSH13-97] - Detector

Sandwich ELISA analysis of human Thrombopoietin matched pair antibodies

Capture: HA725118, Human Thrombopoietin Rabbit mAb [PSH13-96]
Detector: HA725119, Human Thrombopoietin Rabbit mAb [PSH13-97]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725118) 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 Thrombopoietin protein (HA211023) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA725119, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 Thrombopoietin in HepG2 cell culture supernatant.

Capture: HA725118, Human Thrombopoietin Rabbit mAb [PSH13-96]
Detector: HA725119, Human Thrombopoietin Rabbit mAb [PSH13-97]

The concentrations of Thrombopoietin were measured in duplicate and interpolated from the Thrombopoietin standard curve and corrected for sample dilution. Undiluted samples are HepG2 cell culture supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Thrombopoietin concentration was determined to be 47.8 pg/ml in HepG2 cell culture supernatant.