Biotin Conjugated Human DR3/TNFRSF25 Recombinant Rabbit Monoclonal Antibody [PSH16-69] - Detector

Sandwich ELISA analysis of Human DR3/TNFRSF25 matched pair antibodies

Capture: HA723869, Human DR3/TNFRSF25 Rabbit mAb [PSH16-67]
Detector: HA723871, Human DR3/TNFRSF25 Rabbit mAb [PSH16-69]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723869) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/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 DR3/TNFRSF25 protein (HA211322) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723871, 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 DR3/TNFRSF25 in TF-1 and HUVEC extract samples based on a 1,000 µg/ml extract load.

Capture: HA723869, Human DR3/TNFRSF25 Rabbit mAb [PSH16-67]
Detector: HA723871, Human DR3/TNFRSF25 Rabbit mAb [PSH16-69]

The concentrations of DR3/TNFRSF25 were measured in duplicates, interpolated from the DR3/TNFRSF25 standard curve and corrected for sample dilution. Undiluted samples are TF-1 extract 25% and HUVEC extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean DR3/TNFRSF25 concentration was determined to be 2,154 pg/ml in TF-1 extract and undetectable in HUVEC extract.

Interpolated concentrations of spiked DR3/TNFRSF25 in human cell culture media samples.

Capture: HA723869, Human DR3/TNFRSF25 Rabbit mAb [PSH16-67]
Detector: HA723871, Human DR3/TNFRSF25 Rabbit mAb [PSH16-69]

The concentrations of DR3/TNFRSF25 were measured in triplicates, interpolated from the DR3/TNFRSF25 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=3).