Human NCAM1 Recombinant Rabbit Monoclonal Antibody [PSH08-07] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human NCAM1 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722946) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 NCAM1 protein (HA210820) starting from 5000 pg/ml to 0 pg/ml and detect antibody (HA722947, 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 NCAM1 in human samples.

Interpolated concentration of native NCAM1 was measured in duplicate at different sample concentrations. Undiluted samples were 1% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean NCAM1 concentration was determined to be 320 ng/mL in human serum.

Interpolated concentrations of native NCAM1 in human cell culture supernatant samples.

Interpolated concentration of native NCAM1 was measured in duplicate at different sample concentrations. Undiluted samples were 50% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean NCAM1 concentration was determined to be 4621 pg/mL in SH-SY5Y cell culture supernatant samples.

Interpolated concentrations of spiked NCAM1 in cell culture media samples.

The concentrations of NCAM1 were measured in duplicates, interpolated from the NCAM1 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).