Biotin Conjugated Brain-derived tau/BD-tau Recombinant Rabbit Monoclonal Antibody [PSH16-47] - Detector

Sandwich ELISA analysis of Human Brain-derived tau/BD-tau matched pair antibodies

Capture: HA723847, Brain-derived tau/BD-tau Rabbit mAb [PSH16-46]
Detector: HA723848, Brain-derived tau/BD-tau Rabbit mAb [PSH10-47]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723847) 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 Brain-derived tau/BD-tau protein (HA210937) starting from 10000 pg/ml to 0 pg/ml and detect antibody (HA723848, 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 Brain-derived Tau in SH-SY5Y and MDA-MB-231 cell extract samples based on a 1000 µg/ml extract load.

Capture: HA723847, Brain-derived tau/BD-tau Rabbit mAb [PSH16-46]
Detector: HA723848, Brain-derived tau/BD-tau Rabbit mAb [PSH10-47]

Interpolated concentration of native Brain-derived Tau was measured in duplicate at different sample concentrations and interpolated from the Brain-derived Tau standard curves. The mean Brain-derived Tau concentration was determined to be 40,370 pg/mL in SH-SY5Y cell extract. There was no detectable signal in MDA-MB-231 cell extract.

Interpolated concentrations of spiked Brain-derived Tau in cell culture media samples.

Capture: HA723847, Brain-derived tau/BD-tau Rabbit mAb [PSH16-46]
Detector: HA723848, Brain-derived tau/BD-tau Rabbit mAb [PSH10-47]

The concentrations of Brain-derived Tau were measured in duplicates, interpolated from the Brain-derived 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).

Coating Ab: HA723847, 5ug/ml
Co-Incubation: 1:2 serially diluted human Brain-derived Tau protein and Big Tau from HuaBio starting from a concentration of 10,000 pg/ml and Detect Ab: HA723848, Biotin, 0.2 μg/ml (30℃, 120rpm, 60min).
SA-HRP: 1:20,000 (50ng/ml) (30℃, 120rpm, 30min).