Biotin Conjugated Human/Mouse/Rat Synapsin I Recombinant Rabbit Monoclonal Antibody [PSH18-41] - Detector

Sandwich ELISA analysis of human Synapsin 1 matched pair antibodies

Capture: HA723993, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-40]
Detector: HA723994, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-41]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723993) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 Synapsin I protein (HA211359) starting from 48,000 pg/ml to 0 pg/ml and detect antibody (HA723994, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 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 Synapsin-1 in SH-SY5Y cell extract, mouse brain and mouse lung tissue extract samples based on a 1000 µg/ml extract load.

Capture: HA723993, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-40]
Detector: HA723994, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-41]

Interpolated concentration of native Synapsin-1 was measured in duplicate at different sample concentrations and interpolated from the Synapsin-1 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Synapsin-1 concentration was determined to be 7.428 ug/mL in mouse brain tissue extract and 110 ng/mL in SH-SY5Y cell extract. There was no detectable signal in mouse lung tissue extract.

Interpolated concentrations of native Synapsin-1 in rat brain tissue extract samples based on a 1000 µg/ml extract load.

Capture: HA723993, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-40]
Detector: HA723994, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-41]

Interpolated concentration of native Synapsin-1 was measured in duplicate at different sample concentrations and interpolated from the Synapsin-1 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Synapsin-1 concentration was determined to be 6.329 ug/mL in rat brain tissue extract.

Interpolated concentrations of spiked Synapsin-1 in cell culture media samples.

Capture: HA723993, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-40]
Detector: HA723994, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-41]

The concentrations of Synapsin-1 were measured in duplicates, interpolated from the Synapsin-1 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Sandwich ELISA analysis of human Synapsin 1 matched pair antibodies

Capture: HA723993, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-40]
Detector: HA723994, Human/Mouse/Rat Synapsin I Rabbit mAb [PSH18-41]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723993) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 Synapsin I (HA211359) or Human Synapsin 2 protein starting from 48,000 pg/ml to 0 pg/ml and detect antibody (HA723994, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 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.