Biotin Conjugated Human CD31 Recombinant Rabbit Monoclonal Antibody [PSH07-83] - Detector

Sandwich ELISA analysis of Human CD31 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA722906) 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 CD31 protein (HA210891) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA722908B, 0.1 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 CD31in human serum samples.

The concentrations of CD31 were interpolated from the CD31 standard curve and corrected for sample dilution. Undiluted samples are human serum 20%. The mean CD31 concentration was determined to be 39.7 ng/mL in human serum.

Interpolated concentrations of native CD31 in THP-1 and HeLa extract samples based on a 1,000 µg/ml extract load.

The concentrations of CD31 were measured in duplicates, interpolated from the CD31 standard curve and corrected for sample dilution. Undiluted samples are THP-1 extract 3% and HeLa extract 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD31 concentration was determined to be 68.19 ng/ml in THP-1 extract and undetectable in HeLa extract.

Interpolated concentrations of spiked CD31 in human cell culture media samples.

The concentrations of CD31 were measured in duplicates, interpolated from the CD31 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).