Biotin Conjugated Human IKK gamma Recombinant Rabbit Monoclonal Antibody [PSH21-06] - Detector

Sandwich ELISA analysis of Human IKK gamma matched pair antibodies

Capture: HA725327, Human IKK gamma Rabbit mAb [PSH21-05]
Detector: HA725328, Human IKK gamma Rabbit mAb [PSH21-06]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725327) 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 0.2% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human IKK gamma protein (HA211330) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA725328, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 IKK gamma in Hela and Jurkat cell culture lysates.

Capture: HA725327, Human IKK gamma Rabbit mAb [PSH21-05]
Detector: HA725328, Human IKK gamma Rabbit mAb [PSH21-06]

The concentrations of IKK gamma were measured in duplicates, interpolated from the IKK gamma standard curve and corrected for sample dilution. Undiluted HeLa lysate is at 500 μg/ml and undiluted Jurkat lysate is at 250 μg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IKK gamma concentration was determined to be 16.438 ng/ml in Hela cell culture lysate and 29.60ng/ml in Jurkat cell culture lysate.