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Sandwich ELISA analysis of Mouse IL-17A matched pair antibodies
Capture: HA722264, Mouse IL-17A Rabbit mAb [PSH05-35]
Detector: HA722265, Mouse IL-17A Rabbit mAb [PSH05-36]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722264) 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 standard Mouse IL-17A protein (HA210791) starting from 800 pg/ml to 0 pg/ml and detect antibody (HA722265, 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.
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Mouse lung were stimulated with 5 µg/mL LPS and incubated for 6 days.
Capture: HA722264, Mouse IL-17A Rabbit mAb [PSH05-35]
Detector: HA722265, Mouse IL-17A Rabbit mAb [PSH05-36]
The concentrations of IL17A were interpolated from the IL17A standard curves and corrected for sample dilution.Undiluted samples are LPS stimulated mouse lung supernatants 50% and unstimulated mouse lung supernatants 50%. The mean IL17A concentration was determined to be 621 pg/mL in LPS stimulated mouse lung supernatant and undetectable in the unstimulated mouse lung supernatant.
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Interpolated concentrations of native IL17A in unstimulated or stimulated rat spleen culture supernatant.
Capture: HA722264, Mouse IL-17A Rabbit mAb [PSH05-35]
Detector: HA722265, Mouse IL-17A Rabbit mAb [PSH05-36]
Rat spleen were stimulated with 5 µg/mL Concanavalin A and incubated for 2 days.The concentrations of IL17A were interpolated from the IL17A standard curves and corrected for sample dilution. Undiluted samples are Concanavalin A stimulated rat spleen supernatants 20% and unstimulated rat spleen supernatants 50%. The mean IL17A concentration was determined to be 3,703 pg/mL in Concanavalin A stimulated rat spleen supernatant and undetectable in the unstimulated rat spleen supernatant.
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