Biotin Conjugated Human ADAM17 Recombinant Rabbit Monoclonal Antibody [PSH28-42] - Detector

Sandwich ELISA analysis of Human ADAM17 protein matched pair antibodies

Capture: HA725421, Human ADAM17 Rabbit mAb [PSH28-41]
Detector: HA725422, Human ADAM17 Rabbit mAb [PSH28-42]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725421) 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 ADAM17 protein (HA211625) starting from 8,000 pg/mL to 0 pg/mL and detect antibody (HA725422, Biotin, 0.5 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 ADAM17 in THP-1 cell extract and A549 cell extract

Capture: HA725421, Human ADAM17 Rabbit mAb [PSH28-41]
Detector: HA725422, Human ADAM17 Rabbit mAb [PSH28-42]

The concentrations of ADAM17 were measured in duplicates, interpolated from the ADAM17 standard curve and corrected for sample dilution. Undiluted samples are THP-1 cell extract 2 mg/mL and A549 cell extract 2 mg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ADAM17 concentration was determined to be 42.26 ng/mL in THP-1 cell extract and 43.92 ng/mL in A549 cell extract.