Images
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Sandwich ELISA analysis of Human Tau (phospho T217) matched pair antibodies
Capture: HA723841, Phospho-Tau (T217) Rabbit mAb [PSH16-41]
Detector: HA723209, Phospho-Tau (T217) Rabbit mAb [PSH10-42]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723841) 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 Phospho-Tau (T217) protein (HA211033) starting from 50000 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.
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Interpolated concentrations of Tau (phospho T217) in
293T+Human Tau441 2N4R and 293T+OE-Vector Cell Extract samples based on a 1000 µg/ml extract load. 293T+Human Tau441 2N4R represented the overexpression of Tau (phospho T217) protein. 293T+OE-Vector represented the non-expression of Tau (phospho T217) protein.
Capture: HA723841, Phospho-Tau (T217) Rabbit mAb [PSH16-41]
Detector: HA723209, Phospho-Tau (T217) Rabbit mAb [PSH10-42]
Interpolated concentration of Tau (phospho T217) was measured in duplicate at different sample concentrations and interpolated from the Tau (phospho T217) standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Tau (phospho T217) concentration was determined to be 18 ug/mL in 293T+Human Tau441 2N4R cell extract. There was no detectable signal in 293T+OE-Vector cell extract.
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Interpolated concentrations of Tau (phospho T217) in mouse brain tissue extract samples based on a 1000 µg/ml extract load.
Capture: HA723841, Phospho-Tau (T217) Rabbit mAb [PSH16-41]
Detector: HA723209, Phospho-Tau (T217) Rabbit mAb [PSH10-42]
Interpolated concentration of Tau (phospho T217) was measured in duplicate at different sample concentrations and interpolated from the Tau (phospho T217) standard curves. The mean Tau (phospho T217) concentration was determined to be 945,192 pg/mL in mouse brain tissue extract.
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Coating Ab: HA723841, 5ug/mL
Co-Incubation: 1:2 serially diluted human Tau (phospho T217) and Tau (Non-phospho T217) protein from HuaBio starting from a concentration of 50000 pg/ml and Detect Ab: (HA723209, biotin, 0.2 ug/ml) (30℃, 120 rpm, 60min).
SA-HRP:1:20000(50ng/ml) (30°c, 120rpm, 30min).
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Interpolated concentrations of spiked Tau (phospho T217) in cell culture media samples.
Capture: HA723841, Phospho-Tau (T217) Rabbit mAb [PSH16-41]
Detector: HA723209, Phospho-Tau (T217) Rabbit mAb [PSH10-42]
The concentrations of Tau (phospho T217) were measured in duplicates, interpolated from the Tau (phospho T217) 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).
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