Human Mesothelin Recombinant Rabbit Monoclonal Antibody [PSH02-30] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human Mesothelin matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723287) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Mesothelin protein (HA210521) starting from 20000 pg/ml to 0 pg/ml and detect antibody (HA723288, 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 Mesothelin in human cell culture supernatant samples.

Interpolated concentration of native Mesothelin was measured in duplicate at different sample concentrations and interpolated from the Mesothelin standard curves. Undiluted samples were 100% cell supernatant. The mean Mesothelin concentration was determined to be 11,523 pg/mL in neat Hela cell supernanant. There was no detectable signal in PC-3 cell supernatant.

Interpolated concentrations of spiked Mesothelin in cell culture media samples.

The concentrations of Mesothelin were measured in duplicates, interpolated from the Mesothelin standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%.