Biotin Conjugated Human DKK1 Recombinant Rabbit Monoclonal Antibody [PSH21-08] - Detector

Sandwich ELISA analysis of Human DKK1 matched pair antibodies

Capture: HA725241, Human DKK1 Rabbit mAb [PSH21-07]
Detector: HA725242, Human DKK1 Rabbit mAb [PSH21-08]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725241) 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 DKK1 protein (HA211040). starting from 6,000 pg/ml to 0 pg/ml and detect antibody (HA725242, 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 DKK1 in HT-29 supernatant samples.

Capture: HA725241, Human DKK1 Rabbit mAb [PSH21-07]
Detector: HA725242, Human DKK1 Rabbit mAb [PSH21-08]

The concentrations of DKK1 were measured in duplicates, interpolated from the DKK1 standard curve and corrected for sample dilution. Undiluted samples are HT-29 supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean DKK1 concentration was determined to be 30430 pg/ml in HT-29 supernatant.