Biotin Conjugated SNAP25 Recombinant Rabbit Monoclonal Antibody [PSH17-66] - Detector

Sandwich ELISA analysis of Human SNAP25 matched pair antibodies

Capture: HA725227, SNAP25 Rabbit mAb [PSH17-65]
Detector: HA725228, SNAP25 Rabbit mAb [PSH17-66]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725227) 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 SNAP25 protein (HA211155) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA725228, 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 SNAP25 in SH-SY5Y and PANC-1 extract samples based on a 1,000 μg/ml extract load.

Capture: HA725227, SNAP25 Rabbit mAb [PSH17-65]
Detector: HA725228, SNAP25 Rabbit mAb [PSH17-66]

The concentrations of SNAP25 were measured in duplicates, interpolated from the SNAP25 standard curve and corrected for sample dilution. Undiluted samples are SH-SY5Y extract 13% and PANC-1 extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean SNAL25 concentration was determined to be 2,561 pg/ml in SH-SY5Y extract and undetectable in PANC-1 extract.