Human CD85d Recombinant Rabbit Monoclonal Antibody [PSH05-61] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human CD85d matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722381) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 standard Human CD85d protein (HA210643) starting from 15000 pg/ml to 0 pg/ml and detect antibody (HA722382, 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 CD85d in human serum samples.

The concentrations of CD85d were interpolated from the CD85d standard curves and corrected for sample dilution. Undiluted samples are serum 100%. The mean CD85d concentration was determined to be 4448 pg/mL in human serum.

Interpolated concentrations of spiked CD85d in human cell culture media samples.

The concentrations of CD85d were measured in duplicates, interpolated from the CD85d standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).