Biotin Conjugated Human Leptin Receptor Recombinant Rabbit Monoclonal Antibody [PSH14-72] - Detector

Sandwich ELISA analysis of human Leptin Receptor matched pair antibodies

Capture: HA725058, Human Leptin Receptor Rabbit mAb [PSH14-71]
Detector: HA725059, Human Leptin Receptor Rabbit mAb [PSH14-72]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725058) diluted in carbonate/bicarbonate buffer, at a concentration of5µg/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 Leptin Receptor protein (HA211148) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA725059, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 Leptin Receptor in K562 and HepG2 extract samples based on a 1,000 µg/ml extract load.

Capture: HA725058, Human Leptin Receptor Rabbit mAb [PSH14-71]
Detector: HA725059, Human Leptin Receptor Rabbit mAb [PSH14-72]

The concentrations of Leptin Receptor were measured in duplicates, interpolated from the Leptin Receptor standard curve and corrected for sample dilution. Undiluted samples are K562 extract 50% and HepG2 extract 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Leptin Receptor concentration was determined to be 3,255 pg/ml in K562 extract and undetectable in HepG2 extract.