Biotin Conjugated Human CXCL9 Recombinant Rabbit Monoclonal Antibody [PSH12-35] - Detector

Sandwich ELISA analysis of human CXCL9 matched pair antibodies

Capture: HA725004, Human CXCL9 Rabbit mAb [PSH12-34]
Detector: HA725005, Human CXCL9 Rabbit mAb [PSH12-35]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725004) 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 CXCL9 protein (HA210774) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA725005, 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 CXCL9 in THP-1 cell culture supernatant.

Capture: HA725004, Human CXCL9 Rabbit mAb [PSH12-34]
Detector: HA725005, Human CXCL9 Rabbit mAb [PSH12-35]

THP-1 cells were stimulated with 200 ng/ml IFN-γ and 50ng/ml LPS or vehicle control and incubated for 24 hours. The concentrations of CXCL9 measured in duplicate and interpolated from the CXCL9 standard curve and corrected for sample dilution. Undiluted samples are as follows: unstimulated 100% and stimulated 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL9 concentration was determined to be 841.6 pg/ml in IFN-γ and LPS stimulated THP-1 cell culture supernatant and undetectable in the unstimulated THP-1 control.