Human RAGE Recombinant Rabbit Monoclonal Antibody [PSH20-64] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human AGER matched pair antibodies

Capture: HA725289, Human RAGE Rabbit mAb [PSH20-63]
Detector: HA725290, Human RAGE Rabbit mAb [PSH20-64]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725289) 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 RAGE protein (HA211263) starting from10,000 pg/ml to 0 pg/ml and detect antibody (HA725290, 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 AGER in human serum samples.

Capture: HA725289, Human RAGE Rabbit mAb [PSH20-63]
Detector: HA725290, Human RAGE Rabbit mAb [PSH20-64]

The concentrations of AGER were measured in duplicates, interpolated from the AGER standard curve and corrected for sample dilution. Undiluted samples are human serum 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AGER concentration was determined to be 1560 pg/ml in human serum.