Biotin Conjugated Human tPA Recombinant Rabbit Monoclonal Antibody [PSH13-11] - Detector

Sandwich ELISA analysis of Human tPA matched pair antibodies

Capture: HA725115, Human tPA Rabbit mAb [PSH13-10]
Detector: HA725116, Human tPA Rabbit mAb [PSH13-11]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725115) 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 tPA protein (HA211026) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA725116, 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 tPA in HUVEC cell supernatant

Capture: HA725115, Human tPA Rabbit mAb [PSH13-10]
Detector: HA725116, Human tPA Rabbit mAb [PSH13-11]

Interpolated concentration of native tPA was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean tPA concentration was determined to be 17124 pg/mL in HUVEC cell culture supernatant.