Biotin Conjugated Human Acetylcholinesterase Recombinant Rabbit Monoclonal Antibody [PSH17-03] - Detector

Sandwich ELISA analysis of Human Acetylcholinesterase matched pair antibodies

Capture: HA725223, Human Acetylcholinesterase Rabbit mAb [PSH17-02]
Detector: HA725224, Human Acetylcholinesterasen Rabbit mAb [PSH17-03]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725223) 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 Acetylcholinesterase protein (HA211178) starting from 25,000 pg/ml to 0 pg/ml and detect antibody (HA725224, 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 8 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 ACHE in SH-SY5Y extract.

Capture: HA725223, Human Acetylcholinesterase Rabbit mAb [PSH17-02]
Detector: HA725224, Human Acetylcholinesterasen Rabbit mAb [PSH17-03]

The concentrations of ACHE were measured in duplicates, interpolated from the ACHE standard curve and corrected for sample dilution. Undiluted samples are SH-SY5Y extract 1000 ug/ml. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean ACHE concentration was determined to be 1268 pg/ml in SH-SY5Y extract.