Human Alpha-Synuclein Recombinant Rabbit Monoclonal Antibody [JB42-33] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human Alpha-Synuclein matched pair antibodies

Capture: HA723467, Human Alpha-Synuclein Rabbit mAb [JB42-33]
Detector: HA723468, Human Alpha-Synuclein Rabbit mAb [PSH12-56]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723467) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 Human Alpha-synuclein protein (HA211150) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA723468, 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 alpha-Synuclein in SH-SY5Y extract samples based on a 1,000 µg/ml extract load.

Capture: HA723467, Human Alpha-Synuclein Rabbit mAb [JB42-33]
Detector: HA723468, Human Alpha-Synuclein Rabbit mAb [PSH12-56]

The concentrations of alpha-Synuclein were measured in duplicates, interpolated from the alpha-Synuclein standard curve and corrected for sample dilution. Undiluted samples are SH-SY5Y extract 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean alpha-Synuclein concentration was determined to be 2,195 pg/ml in SH-SY5Y extract.

Interpolated concentrations of native alpha-Synuclein in human serum samples.

Capture: HA723467, Human Alpha-Synuclein Rabbit mAb [JB42-33]
Detector: HA723468, Human Alpha-Synuclein Rabbit mAb [PSH12-56]

The concentrations of alpha-Synuclein were measured in duplicates, interpolated from the alpha-Synuclein standard curve and corrected for sample dilution. Undiluted samples are human serum 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean alpha-Synuclein concentration was determined to be 21.2 ng/ml in human serum.

Interpolated concentrations of spiked alpha-Synuclein in human cell culture media samples.

Capture: HA723467, Human Alpha-Synuclein Rabbit mAb [JB42-33]
Detector: HA723468, Human Alpha-Synuclein Rabbit mAb [PSH12-56]

The concentrations of alpha-Synuclein were measured in duplicates, interpolated from the alpha-Synuclein standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).