Human IL-22 Recombinant Rabbit Monoclonal Antibody [PSH03-12] - BSA and Azide free (Detector)

Sandwich ELISA analysis of human IL-22 matched pair antibodies

Capture: HA721939, Mouse CRP Rabbit mAb [PSH03-11]
Detector: HA721940, Mouse CRP Rabbit mAb [PSH03-12]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA721939) 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 IL-22 protein (HA210691) starting from 2000 pg/ml to 0 pg/ml and detect antibody (HA721940, 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.

Conditioned media was harvested after 48 hours. IL-22 was measured in 100% unstimulated and PHA stimulated PBMC supernatant.The concentrations of IL-22 were interpolated from the IL-22 standard curves and corrected for sample dilution.The mean IL-22 concentration was determined to be 79.7 pg/mL in PHA stimulated PBMC supernatant and there was no detectable signal in unstimulated supernatant.