Biotin Conjugated Human DLL4 Recombinant Rabbit Monoclonal Antibody [PSH29-27] - Detector

Sandwich ELISA analysis of Human DLL4 protein matched pair antibodies

Capture: HA724855, Human DLL4 Rabbit mAb [PSH29-26]
Detector: HA724856, Human DLL4 Rabbit mAb [PSH29-27]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724855) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 DLL4 protein (HA211588) starting from 20,000 pg/mL to 0 pg/mL and detect antibody (HA724856, Biotin, 0.1 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 DLL4 in HUVEC and HeLa extract samples based on a 1,000 µg/mL extract load.

Capture: HA724855, Human DLL4 Rabbit mAb [PSH29-26]
Detector: HA724856, Human DLL4 Rabbit mAb [PSH29-27]

Interpolated concentration of native DLL4 was measured in duplicate at different sample concentrations and interpolated from the DLL4 standard curves. The mean DLL4 concentration was determined to be 1,250 pg/mL in HUVEC cell extract. There was no detectable signal in HeLa cell extract.

Interpolated concentrations of spiked DLL4 in cell culture media samples.

Capture: HA724855, Human DLL4 Rabbit mAb [PSH29-26]
Detector: HA724856, Human DLL4 Rabbit mAb [PSH29-27]

The concentrations of DLL4 were measured in duplicates, interpolated from the DLL4 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).