Biotin Conjugated Human IL-1 beta Recombinant Rabbit Monoclonal Antibody [PSH16-86] - Detector

Sandwich ELISA analysis of human IL1β matched pair antibodies

Capture: HA723881, Human IL-1 beta Rabbit mAb [PSH16-85]
Detector: HA723882, Human IL-1 beta Rabbit mAb [PSH16-86]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723881) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 IL-1 beta protein (HA210576) starting from 1000 pg/ml to 0 pg/ml and detect antibody (HA723882, 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 human IL1β matched pair antibodies

Capture: HA723881, Human IL-1 beta Rabbit mAb [PSH16-87]
Detector: HA723882, Human IL-1 beta Rabbit mAb [PSH16-86]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723883) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 IL-1 beta protein (HA210576) starting from 1000 pg/ml to 0 pg/ml and detect antibody (HA723882, 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 IL-1 beta in human THP-1 supernatant treated or untreated with LPS for 48 hours.

Capture: HA723881, Human IL-1 beta Rabbit mAb [PSH16-85]
Detector: HA723882, Human IL-1 beta Rabbit mAb [PSH16-86]

Interpolated concentration of native IL-1 beta was measured in duplicate at different sample concentrations and interpolated from the IL-1 beta standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1 beta concentration was determined to be 187 pg/mL in neat THP-1 treated supernatant and undetectable in untreated THP-1 supernatant.

Interpolated concentrations of native IL-1 beta in human THP-1 supernatant treated or untreated with LPS for 48 hours.

Capture: HA723881, Human IL-1 beta Rabbit mAb [PSH16-87]
Detector: HA723882, Human IL-1 beta Rabbit mAb [PSH16-86]

Interpolated concentration of native IL-1 beta was measured in duplicate at different sample concentrations and interpolated from the IL-1 beta standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean IL-1 beta concentration was determined to be 293 pg/mL in neat THP-1 treated supernatant and undetectable in untreated THP-1 supernatant.

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

Capture: HA723881, Human IL-1 beta Rabbit mAb [PSH16-85]
Detector: HA723882, Human IL-1 beta Rabbit mAb [PSH16-86]

The concentrations of IL-1 beta were measured in duplicates, interpolated from the IL-1 beta standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

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

Capture: HA723881, Human IL-1 beta Rabbit mAb [PSH16-87]
Detector: HA723882, Human IL-1 beta Rabbit mAb [PSH16-86]

The concentrations of IL-1 beta were measured in duplicates, interpolated from the IL-1 beta standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).