Human ICAM3 Recombinant Rabbit Monoclonal Antibody [PSH20-69] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human ICAM3 matched pair antibodies

Capture: HA724167, Human ICAM3 Rabbit mAb [PSH20-69]
Detector: HA724168, Human ICAM3 Rabbit mAb [PSH20-70]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724167) 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 0.2% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human ICAM3 protein (HA211145) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA724168, 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-3 in human serum samples.

Capture: HA724167, Human ICAM3 Rabbit mAb [PSH20-69]
Detector: HA724168, Human ICAM3 Rabbit mAb [PSH20-70]

The concentrations of ICAM-3 were measured in duplicates, interpolated from the ICAM-3 standard curve and corrected for sample dilution. Undiluted samples are human serum 2%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM-3 concentration was determined to be 216 ng/ml in human serum.

Interpolated concentrations of spiked ICAM-3 in human cell culture media samples.

Capture: HA724167, Human ICAM3 Rabbit mAb [PSH20-69]
Detector: HA724168, Human ICAM3 Rabbit mAb [PSH20-70]

The concentrations of ICAM-3 were measured in duplicates, interpolated from the ICAM-3 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).