Human IDO1 Recombinant Rabbit Monoclonal Antibody [PSH09-86] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human IDO matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723154) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 IDO1 protein (HA210938) starting from 10000 pg/ml to 0 pg/ml and detect antibody (HA723155, 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 IDO in IFN-r stimulated Hela cell extracts based on a 1000 ug/ml extract load.

Interpolated concentration of native IDO was measured in duplicate at different sample concentrations and interpolated from the IDO standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IDO concentration was determined to be 158,961 pg/mL in IFN-r stimulated Hela cell extracts. IDO was not detected in naive HeLa cells with a 1000 ug/mL extract load.

Interpolated concentrations of spiked IDO in cell culture media samples.

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