Tau Recombinant Rabbit Monoclonal Antibody [PSH16-48] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human Tau matched pair antibodies

Capture: HA723850, Tau Rabbit mAb [PSH16-48]
Detector: HA723209, Tau Rabbit mAb [PSH10-42]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723850) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Tau protein (HA210937) starting from 10000 pg/ml to 0 pg/ml and detect antibody (HA723209, 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.

Sandwich ELISA analysis of human Tau matched pair antibodies

Capture: HA723850, Tau Rabbit mAb [PSH16-48]
Detector: HA723209, Tau Rabbit mAb [PSH10-42]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723850) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Tau protein (HA211246) starting from 10000 pg/ml to 0 pg/ml and detect antibody (HA723209, 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 Tau in SH-SY5Y and mouse cerebellum extract samples based on a 1000 µg/ml extract load.

Capture: HA723850, Tau Rabbit mAb [PSH16-48]
Detector: HA723209, Tau Rabbit mAb [PSH10-42]

Interpolated concentration of native Tau was measured in duplicate at different sample concentrations and interpolated from the Tau standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Tau concentration was determined to be 14,861 pg/mL in SH-SY5Y cell extract and 3,257 pg/mL in mouse cerebellum tissue extract.

Interpolated concentrations of spiked Tau in cell culture media samples.

Capture: HA723850, Tau Rabbit mAb [PSH16-48]
Detector: HA723209, Tau Rabbit mAb [PSH10-42]

The concentrations of Tau were measured in duplicates, interpolated from the Tau 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 Tau matched pair antibodies

Capture: HA723850, Tau Rabbit mAb [PSH16-48]
Detector: HA723851, Tau Rabbit mAb [PSH16-49]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723850) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Tau protein (HA210937) starting from 10000 pg/ml to 0 pg/ml and detect antibody (HA723851, 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 Tau (4R Isoforms) in SH-SY5Y extract samples based on a 1000 µg/ml extract load.

Capture: HA723850, Tau Rabbit mAb [PSH16-48]
Detector: HA723851, Tau Rabbit mAb [PSH16-49]

Interpolated concentration of native Tau (4R Isoforms) was measured in duplicate at different sample concentrations and interpolated from the Tau (4R Isoforms) standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Tau (4R Isoforms) concentration was determined to be 41,927 pg/mL in SH-SY5Y cell extract.

Interpolated concentrations of native Tau (4R Isoforms) in human cell culture supernatant samples.

Capture: HA723850, Tau Rabbit mAb [PSH16-48]
Detector: HA723851, Tau Rabbit mAb [PSH16-49]

Interpolated concentration of native Tau (4R Isoforms) was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Tau (4R Isoforms) concentration was determined to be 2,372 pg/mL in SH-SY5Y cell supernatant. There was no detectable signal in MDA-MB-231 cell culture supernatant.

Interpolated concentrations of spiked Tau (4R Isoforms) in cell culture media samples.

Capture: HA723850, Tau Rabbit mAb [PSH16-48]
Detector: HA723851, Tau Rabbit mAb [PSH16-49]

The concentrations of Tau (4R Isoforms) were measured in duplicates, interpolated from the Tau (4R Isoforms) 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).