Biotin Conjugated Human LAMC2 Recombinant Rabbit Monoclonal Antibody [PSH15-15] - Detector

Sandwich ELISA analysis of human LAMC2 matched pair antibodies

Capture: HA723717, Human LAMC2 Rabbit mAb [PSH15-14]
Detector: HA723718, Human LAMC2 Rabbit mAb [PSH15-15]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723717) 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 LAMC2 protein (HA211219) starting from 6,000 pg/ml to 0 pg/ml and detect antibody (HA723718, 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 LAMC2 in human samples.

Capture: HA723717, Human LAMC2 Rabbit mAb [PSH15-14]
Detector: HA723718, Human LAMC2 Rabbit mAb [PSH15-15]

Interpolated concentration of native LAMC2 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean LAMC2 concentration was determined to be 105,837 pg/mL in A431 and 1,733 pg/ml in A549 cell culture supernatant. There was no detectable signal in HepG2 cell culture supernatant.

Interpolated concentrations of spiked LAMC2 in cell culture media samples.

Capture: HA723717, Human LAMC2 Rabbit mAb [PSH15-14]
Detector: HA723718, Human LAMC2 Rabbit mAb [PSH15-15]

The concentrations of LAMC2 were measured in duplicates, interpolated from the LAMC2 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).