Biotin Conjugated Human CD83 Recombinant Rabbit Monoclonal Antibody [PSH20-09] - Detector

Sandwich ELISA analysis of Human CD83 matched pair antibodies

Capture: HA725305, Human CD83 Rabbit mAb [PSH20-08]
Detector: HA725306, Human CD83 Rabbit mAb [PSH20-09]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725305) 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 CD83 protein (HA211203) starting from 7,000 pg/ml to 0 pg/ml and detect antibody (HA725306, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 CD83 in Raji samples.

Capture: HA725305, Human CD83 Rabbit mAb [PSH20-08]
Detector: HA725306, Human CD83 Rabbit mAb [PSH20-09]

The concentrations of CD83 were measured in duplicates, interpolated from the CD83 standard curve and corrected for sample dilution. Undiluted samples are Raji supernatant 100% and THP-1 supernatant. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD83 concentration was determined to be 148 pg/ml in Raji.