Mouse IL-1RA Recombinant Rabbit Monoclonal Antibody [PSH14-99] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Mouse IL-1RA matched pair antibodies

Capture: HA723697, Mouse IL-1RA Rabbit mAb [PSH14-98]
Detector: HA723698, Mouse IL-1RA Rabbit mAb [PSH14-99]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723697) 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 IL-1RA protein (HA211041) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723698, 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.

Sandwich ELISA analysis of Mouse IL-1RA matched pair antibodies

Capture: HA723700, Mouse IL-1RA Rabbit mAb [PSH15-00]
Detector: HA723698, Mouse IL-1RA Rabbit mAb [PSH14-99]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723700) 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 IL-1RA protein (HA211041) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723698, 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 c in J774A.1 cell
culture supernatant untreated or treated with LPS and IFNγ for 24h.

Capture: HA723697, Mouse IL-1RA Rabbit mAb [PSH14-98]
Detector: HA723698, Mouse IL-1RA Rabbit mAb [PSH14-99]

Interpolated concentration of native IL-1RA was measured in duplicate at different sample concentrations and interpolated from the IL-1RA standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1RA concentration was determined to be 131 pg/ml in J774A.1 untreated cell culture supernatant,10634 pg/ml in J774A.1 treated cell culture supernatant.

Interpolated concentrations of native IL-1RA in J774A.1 cell
culture supernatant untreated or treated with LPS and IFNγ for 24h.

Capture: HA723700, Mouse IL-1RA Rabbit mAb [PSH15-00]
Detector: HA723698, Mouse IL-1RA Rabbit mAb [PSH14-99]

Interpolated concentration of native IL-1RA was measured in duplicate at different sample concentrations and interpolated from the IL-1RA standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1RA concentration was determined to be 1848 pg/ml in J774A.1 untreated cell culture supernatant, 47377 pg/ml in J774A.1 treated cell culture supernatant.

Interpolated concentrations of spiked IL-1RA in cell culture media samples.

Capture: HA723697, Mouse IL-1RA Rabbit mAb [PSH14-98]
Detector: HA723698, Mouse IL-1RA Rabbit mAb [PSH14-99]

The concentrations of IL-1RA were measured in duplicates, interpolated from the IL-1RA 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 spiked IL-1RA in cell culture media samples.

Capture: HA723700, Mouse IL-1RA Rabbit mAb [PSH15-00]
Detector: HA723698, Mouse IL-1RA Rabbit mAb [PSH14-99]

The concentrations of IL-1RA were measured in duplicates, interpolated from the IL-1RA 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).