Biotin Conjugated Human IL-6R Recombinant Rabbit Monoclonal Antibody [PSH15-39] - Detector

Sandwich ELISA analysis of Human IL-6R matched pair antibodies

Capture: HA723743, Human IL-6R Rabbit mAb [PSH15-38]
Detector: HA723744, Human IL-6R Rabbit mAb [PSH15-39]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723743) 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 IL-6R protein (HA210640) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA723744, 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 IL-6R in THP-1 and Daudi cell culture supernatant .

Capture: HA723743, Human IL-6R Rabbit mAb [PSH15-38]
Detector: HA723744, Human IL-6R Rabbit mAb [PSH15-39]

Interpolated concentration of native IL-6R was measured in duplicate at different sample concentrations and interpolated from the IL-6R standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-6R concentration was determined to be 3824pg/mL in THP-1 cell culture supernatan, undetectable in Daudi cell culture supernatan.

Interpolated concentrations of spiked IL-6R in cell culture media samples.

Capture: HA723743, Human IL-6R Rabbit mAb [PSH15-38]
Detector: HA723744, Human IL-6R Rabbit mAb [PSH15-39]

The concentrations of IL-6R were measured in duplicates, interpolated from the IL-6R standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).