Mouse IFN gamma Recombinant Rabbit Monoclonal Antibody [PSH14-81] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Mouse IFN gamma matched pair antibodies

Capture: HA723676, Mouse IFN gamma Rabbit mAb [PSH14-80]
Detector: HA723677, Mouse IFN gamma Rabbit mAb [PSH14-81]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723676) 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 Mouse IFN gamma protein (HA210974) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723677, 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 IFN gamma in mouse spleen samples treated with or without 5 µg/ml LPS for 6 days.

Capture: HA723676, Mouse IFN gamma Rabbit mAb [PSH14-80]
Detector: HA723677, Mouse IFN gamma Rabbit mAb [PSH14-81]

The concentrations of IFN gamma measured in duplicate and interpolated from the IFN gamma standard curve and corrected for sample dilution. Undiluted samples are as follows: unstimulated 100% and stimulated 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IFN gamma concentration was determined to be 1,481 pg/ml in LPS stimulated mouse spleen supernatant and undetectable in the unstimulated mouse spleen supernatant.

Interpolated concentrations of spiked IFN gamma in mouse cell culture media samples.

Capture: HA723676, Mouse IFN gamma Rabbit mAb [PSH14-80]
Detector: HA723677, Mouse IFN gamma Rabbit mAb [PSH14-81]

The concentrations of IFN gamma were measured in duplicates, interpolated from the IFN gamma 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).

Interpolated concentrations of native IFN gamma in male / female mouse spleen samples treated with or without 400 µg/ml LPS for 6 hours.

Capture: HA723676, Mouse IFN gamma Rabbit mAb [PSH14-80]
Detector: HA723677, Mouse IFN gamma Rabbit mAb [PSH14-81]

Interpolated concentration of native IFN gamma was measured in duplicate at different sample concentrations and interpolated from the IFN gamma standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IFN gamma concentration were determined to be 2,445 pg/mL in LPS stimulated male mouse spleen supernatant, 2,102 pg/ml in LPS stimulated female mouse spleen supernatant and 704 pg/ml in unstimulated male mouse spleen supernatant.