Mouse CXCL9 Recombinant Rabbit Monoclonal Antibody [PSH12-63] - BSA and Azide free (Detector)

Sandwich ELISA analysis of mouse CXCL9 matched pair antibodies

Capture: HA725022, Mouse CXCL9 Rabbit mAb [PSH12-62]
Detector: HA725023, Mouse CXCL9 Rabbit mAb [PSH12-63]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725022) 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 Mouse CXCL9 protein (HA211031) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA725023, 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 unstimulated and stimulated J774A.1 cell culture supernatant.

Capture: HA725022, Mouse CXCL9 Rabbit mAb [PSH12-62]
Detector: HA725023, Mouse CXCL9 Rabbit mAb [PSH12-63]

J774A.1 cells were stimulated with 20 ng/ml recombinant mouse IFN-γ and 1 μg/ml LPS incubated for 1 day. 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 1.6%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL9 concentration was determined to be 17,738 pg/ml in IFN-γ and LPS stimulated J774A.1 cell culture supernatant and undetectable in the unstimulated J774A.1 control.