Biotin Conjugated Rat ICAM1 Recombinant Rabbit Monoclonal Antibody [PSH17-52] - Detector

Sandwich ELISA analysis of Rat ICAM1 matched pair antibodies

Capture: HA723926, Rat ICAM1 Rabbit mAb [PSH17-51]
Detector: HA723927, Rat ICAM1 Rabbit mAb [PSH17-52]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723926) 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 Rat ICAM1 protein (HA211063) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723927, 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 Rat ICAM1 matched pair antibodies

Capture: HA723928, Rat ICAM1 Rabbit mAb [PSH17-53]
Detector: HA723927, Rat ICAM1 Rabbit mAb [PSH17-52]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723928) 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 Rat ICAM1 protein (HA211063) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723927, 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 ICAM1 in rat serum samples.

Capture: HA723926, Rat ICAM1 Rabbit mAb [PSH17-51]
Detector: HA723927, Rat ICAM1 Rabbit mAb [PSH17-52]

The concentrations of ICAM1 were measured in duplicates, interpolated from the ICAM1 standard curve and corrected for sample dilution. Undiluted samples are rat serum 2.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM1 concentration was determined to be 62,341 pg/ml in rat serum.

Interpolated concentrations of native ICAM1 in rat serum samples.

Capture: HA723928, Rat ICAM1 Rabbit mAb [PSH17-53]
Detector: HA723927, Rat ICAM1 Rabbit mAb [PSH17-52]

The concentrations of ICAM1 were measured in duplicates, interpolated from the ICAM1 standard curve and corrected for sample dilution. Undiluted samples are rat serum 5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM1 concentration was determined to be 21,617 pg/ml in rat serum.

Interpolated concentrations of spiked ICAM1 in rat cell culture media samples.

Capture: HA723926, Rat ICAM1 Rabbit mAb [PSH17-51]
Detector: HA723927, Rat ICAM1 Rabbit mAb [PSH17-52]

The concentrations of ICAM1 were measured in duplicates, interpolated from the ICAM1 standard curves and corrected for sample dilution. Diluted samples are as follows: 25% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Interpolated concentrations of spiked ICAM1 in rat cell culture media samples.

Capture: HA723928, Rat ICAM1 Rabbit mAb [PSH17-53]
Detector: HA723927, Rat ICAM1 Rabbit mAb [PSH17-52]

The concentrations of ICAM1 were measured in duplicates, interpolated from the ICAM1 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).