Human BACE1 Recombinant Rabbit Monoclonal Antibody [PSH16-23] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human BACE1 matched pair antibodies

Capture: HA725171, Human BACE1 Rabbit mAb [PSH16-22]
Detector: HA725172, Human BACE1 Rabbit mAb [PSH16-23]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725171) 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 BACE1 protein (HA211177) protein starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA725172, 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 BACE1 in SH-SY5Y samples based on a 1,000µg/mL extract load.

Capture: HA725171, Human BACE1 Rabbit mAb [PSH16-22]
Detector: HA725172, Human BACE1 Rabbit mAb [PSH16-23]

The concentrations of BACE1 were measured in duplicates, interpolated from the BACE1 standard curve and corrected for sample dilution. Undiluted samples are SH-SY5Y extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean human BACE1 concentration was determined to be 7417.89 pg/ml in SH-SY5Y extract.