Biotin Conjugated Mouse CD30 Recombinant Rabbit Monoclonal Antibody [PSH17-55] - Detector

Sandwich ELISA analysis of Mouse CD30 matched pair antibodies

Capture: HA723929, Mouse CD30 Rabbit mAb [PSH17-54]
Detector: HA723930, Mouse CD30 Rabbit mAb [PSH17-55]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723929) 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 Mouse CD30 protein (HA211062) starting from 8000 pg/ml to 0 pg/ml and detect antibody (HA723930, 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 CD30 in mouse spleen cell supernatant treated or untreated with IL-2 and Concanavalin A.

Capture: HA723929, Mouse CD30 Rabbit mAb [PSH17-54]
Detector: HA723930, Mouse CD30 Rabbit mAb [PSH17-55]

Interpolated concentration of native CD30 was measured in duplicate at different sample concentrations and interpolated from the CD30 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD30 concentration was determined to be 9,657 pg/mL in treated mouse spleen cell supernatant. There was no detectable signal in untreated mouse spleen cell supernatant.

Interpolated concentrations of spiked CD30 in cell culture media samples.

Capture: HA723929, Mouse CD30 Rabbit mAb [PSH17-54]
Detector: HA723930, Mouse CD30 Rabbit mAb [PSH17-55]

The concentrations of CD30 were measured in duplicates, interpolated from the Osteopontin CD30 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).