Human VCAM-1 Recombinant Rabbit Monoclonal Antibody [PSH12-47] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human VCAM-1 matched pair antibodies

Capture: HA725064, Human VCAM-1 Rabbit mAb [PSH12-46]
Detector: HA725065, Human VCAM-1 Rabbit mAb [PSH12-47]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725064) 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 VCAM-1 protein (HA210976) protein starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA725065, 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 VCAM-1 in human samples.

Capture: HA725064, Human VCAM-1 Rabbit mAb [PSH12-46]
Detector: HA725065, Human VCAM-1 Rabbit mAb [PSH12-47]

Interpolated concentration of native VCAM-1 was measured in duplicate at different sample concentrations. Undiluted samples were 1% human serum. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean VCAM-1 concentration was determined to be 1,361,999 pg/mL in human serum. There was no detectable signal in SK-OV-3 cell culture supernatant.