Biotin Conjugated Human ICAM2 Recombinant Rabbit Monoclonal Antibody [PSH25-48] - Detector

Sandwich ELISA analysis of Human ICAM2 matched pair antibodies

Capture: HA724538, Human ICAM2 Rabbit mAb [PSH25-47]
Detector: HA724539, Human ICAM2 Rabbit mAb [PSH25-48]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724538) 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 Human ICAM2 protein (HA211314) starting from 5,000 pg/mL to 0 pg/mL and detect antibody (HA724539, 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 ICAM2 in human serum samples, Jurkat and HeLa cell culture supernatant.

Capture: HA724538, Human ICAM2 Rabbit mAb [PSH25-47]
Detector: HA724539, Human ICAM2 Rabbit mAb [PSH25-48]

The concentrations of ICAM2 were measured in duplicates, interpolated from the ICAM2 standard curve and corrected for sample dilution. Undiluted samples are human serum 0.25%, Jurkat cell culture supernatant 50%, Hela cell culture supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ICAM2 concentration was determined to be 269,883 pg/mL in human serum, 1,312 pg/mL in Jurkat cell culture supernatant and undetectable in HeLa cell culture supernatant.

Interpolated concentrations of spiked ICAM2 in human cell culture media samples.

Capture: HA724538, Human ICAM2 Rabbit mAb [PSH25-47]
Detector: HA724539, Human ICAM2 Rabbit mAb [PSH25-48]

The concentrations of ICAM2 were measured in duplicates, interpolated from the ICAM2 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).