Biotin Conjugated Human PCSK9 Recombinant Rabbit Monoclonal Antibody [PSH12-86] - Detector

Sandwich ELISA analysis of Human PCSK9 matched pair antibodies

Capture: HA725124, Human PCSK9 Rabbit mAb [PSH12-85]
Detector: HA725125, Human PCSK9 Rabbit mAb [PSH12-86]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725124) 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 PCSK9 protein (HA210991) starting from 100,000 pg/ml to 0 pg/ml and detect antibody (HA725125, 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 PCSK9 in HepG2 cell culture supernatant and cell extract.

Capture: HA725124, Human PCSK9 Rabbit mAb [PSH12-85]
Detector: HA725125, Human PCSK9 Rabbit mAb [PSH12-86]

The concentrations of PCSK9 were measured in duplicates, interpolated from the PCSK9 standard curve and corrected for sample dilution. Undiluted samples are HepG2 cell culture supernatant 50% and HepG2 cell extract 250 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean PCSK9 concentration was determined to be 9,074 pg/ml in HepG2 cell culture supernatant and 46,405 pg/ml in HepG2 cell extract.