Human CD30 Recombinant Rabbit Monoclonal Antibody [PSH04-82] - BSA and Azide free (Detector)

Sandwich ELISA analysis of human CD30 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA722187) 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 CD30 protein (HA210649) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA722188) (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.

The concentrations of CD30 were interpolated from the CD30 standard curves and corrected for sample dilution. Undiluted samples are HDLM-2 cell culture supernatant 50%. The mean CD30 concentration was determined to be 1,260 pg/ml in HDLM-2 cell culture supernatant.

The concentrations of CD30 were interpolated from the CD30 standard curves and corrected for sample dilution. Undiluted samples are mixed serum from twenty-four volunteers 13%. The mean CD30 concentration was determined to be 11,531 pg/ml in mixed serum from twenty-four volunteers.