Biotin Conjugated Human PAI1 Recombinant Rabbit Monoclonal Antibody [PSH12-18] - Detector

Sandwich ELISA analysis of Human PAI1 matched pair antibodies

Capture: HA723438, Human PAI1 Rabbit mAb [PSH12-20]
Detector: HA723435, Human PAI1 Rabbit mAb [PSH12-18]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723438) 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 PAI1 protein (HA210982) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA723435, 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 PAI1 in human cell culture supernatant samples.

Capture: HA723438, Human PAI1 Rabbit mAb [PSH12-20]
Detector: HA723435, Human PAI1 Rabbit mAb [PSH12-18]

Interpolated concentration of native PAI1 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean PAI1 concentration was determined to be 371000 pg/mL in HUVEC sups, 51000 pg/ml in HepG2 cell culture supernatant.

Interpolated concentrations of spiked PAI1 in cell culture media samples.

Capture: HA723438, Human PAI1 Rabbit mAb [PSH12-20]
Detector: HA723435, Human PAI1 Rabbit mAb [PSH12-18]

The concentrations of PAI1 were measured in duplicates, interpolated from the PAI1 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).