Biotin Conjugated Mouse ICAM1 Recombinant Rabbit Monoclonal Antibody [PSH16-35] - Detector

Sandwich ELISA analysis of Mouse ICAM1 matched pair antibodies

Capture: HA723832, Mouse ICAM1 Rabbit mAb [PSH16-34]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723832) 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 ICAM1 protein (HA211066) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723833, 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 ICAM1 matched pair antibodies

Capture: HA723834, Mouse ICAM1 Rabbit mAb [PSH16-36]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723834) 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 ICAM1 protein (HA211066) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723833, 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 ICAM-1 in mouse serum samples.

Capture: HA723832, Mouse ICAM1 Rabbit mAb [PSH16-34]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

The concentrations of ICAM-1 were measured in duplicates, interpolated from the ICAM-1 standard curve and corrected for sample dilution. Undiluted samples are mouse serum 0.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-1 concentration was determined to be 930.1 ng/ml in mouse serum.

Interpolated concentrations of native ICAM-1 in mouse serum samples.

Capture: HA723834, Mouse ICAM1 Rabbit mAb [PSH16-36]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

The concentrations of ICAM-1 were measured in duplicates, interpolated from the ICAM-1 standard curve and corrected for sample dilution. Undiluted samples are mouse serum 0.125%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-1 concentration was determined to be 1,171 ng/ml in mouse serum.

Interpolated concentrations of native ICAM-1 in mouse LPS and mouse IFN-γ stimulated J774A.1 cell culture supernatant.

Capture: HA723832, Mouse ICAM1 Rabbit mAb [PSH16-34]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

J774A.1 cells were stimulated with 1 µg/ml LPS and 100 ng/ml mouse IFN-γ or vehicle control and incubated for 72 hours. The concentrations of ICAM-1 measured in duplicate and interpolated from the ICAM-1 standard curve and corrected for sample dilution. Undiluted samples are as follows: stimulated 20%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-1 concentration was determined to be 6,529 pg/ml in LPS and mouse IFN-γ stimulated J774A.1 cell culture supernatant.

Interpolated concentrations of native ICAM-1 in mouse LPS and mouse IFN-γ stimulated J774A.1 cell culture supernatant.

Capture: HA723834, Mouse ICAM1 Rabbit mAb [PSH16-36]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

J774A.1 cells were stimulated with 1 µg/ml LPS and 100 ng/ml mouse IFN-γ or vehicle control and incubated for 72 hours. The concentrations of ICAM-1 measured in duplicate and interpolated from the ICAM-1 standard curve and corrected for sample dilution. Undiluted samples are as follows: stimulated 20%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-1 concentration was determined to be 5,372 pg/ml in LPS and mouse IFN-γ stimulated J774A.1 cell culture supernatant.

Interpolated concentrations of spiked ICAM-1 in mouse cell culture media samples.

Capture: HA723832, Mouse ICAM1 Rabbit mAb [PSH16-34]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

The concentrations of ICAM-1 were measured in duplicates, interpolated from the ICAM-1 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 ICAM-1 in mouse cell culture media samples.

Capture: HA723834, Mouse ICAM1 Rabbit mAb [PSH16-36]
Detector: HA723833, Mouse ICAM1 Rabbit mAb [PSH16-35]

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