Human Prolactin Recombinant Rabbit Monoclonal Antibody [PSH03-14] - BSA and Azide free (Detector)

Sandwich ELISA analysis of human Prolactin matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA721941) 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 Prolactin protein starting from 5000 pg/ml to 0 pg/ml and detect antibody (HA721942)-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.

The concentrations of Prolactin were interpolated from the Prolactin standard curves and corrected for sample dilution.The Prolactin concentration was determined to be 6,297 pg/mL in serum 20%, 6,367 pg/ml in serum 10%, 5,913 pg/ml in serum 5%, 7,660 pg/ml in serum 2.5%. The mean Prolactin concentration was determined to be 6,559 pg/ml in mixed serum from twenty volunteers.

The concentrations of Prolactin were interpolated from the Prolactin standard curves and corrected for sample dilution.The mean Prolactin concentration was determined to be 2,995 pg/ml in Daudi extract.