Biotin Conjugated Human Lipocalin-2 Recombinant Rabbit Monoclonal Antibody [PSH13-68] - Detector

Sandwich ELISA analysis of Human Lipocalin-2 matched pair antibodies

Capture: HA725109, Human Lipocalin-2 Rabbit mAb [PSH13-67]
Detector: HA725110, Human Lipocalin-2 Rabbit mAb [PSH13-68]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725109) 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 Lipocalin-2 protein (HA211028) starting from 6,000 pg/ml to 0 pg/ml and detect antibody (HA725110, 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 Lipocalin-2 in human samples.

Capture: HA725109, Human Lipocalin-2 Rabbit mAb [PSH13-67]
Detector: HA725110, Human Lipocalin-2 Rabbit mAb [PSH13-68]

Interpolated concentration of native Lipocalin-2 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Lipocalin-2 concentration was determined to be 269,116 pg/mL in A431 cell culture supernatant and 2,421pg/mL in Human urine.