Images
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Sandwich ELISA analysis of Human Non-p217 Tau matched pair antibodies
Capture: HA723843, Non-Phospho-Tau (T217) Rabbit mAb [PSH16-43]
Detector: HA723209, Non-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 (HA723843) 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 (HA210937) starting from 30000 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 native Non-p217 Tau in rat cerebellum, SH-SY5Y and MDA-MB-231 extract samples based on a 1000 µg/ml extract load.
Capture: HA723843, Non-Phospho-Tau (T217) Rabbit mAb [PSH16-43]
Detector: HA723209, Non-Phospho-Tau (T217) Rabbit mAb [PSH10-42]
Interpolated concentration of native Non-p217 Tau was measured in duplicate at different sample concentrations and interpolated from the Non-p217 Tau standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Non-p217 Tau concentration was determined to be 561,264 pg/mL in rat cerebellum and 66,440 pg/mL in SH-SY5Y cell extract, There was no detectable signal in MDA-MB-231 cell extract.
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Interpolated concentrations of spiked Non-p217 Tau in cell culture media samples.
Capture: HA723843, Non-Phospho-Tau (T217) Rabbit mAb [PSH16-43]
Detector: HA723209, Non-Phospho-Tau (T217) Rabbit mAb [PSH10-42]
The concentrations of Non-p217 Tau were measured in duplicates, interpolated from the Non-p217 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).
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