Human CD200 Recombinant Rabbit Monoclonal Antibody [PSH05-59] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human CD200 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722379) 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 CD200 protein (HA210665) starting from 2,500 pg/ml to 0 pg/ml and detect antibody (HA722380, 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 CD200 in human serum samples.

The concentrations of CD200 were interpolated from the CD200 standard curves and corrected for sample dilution. Undiluted samples are human serum 100%. The mean CD200 concentration was determined to be 652.9 pg/ml in human serum.

Interpolated concentrations of native CD200 in SK-Mel-28 and NCI-H226 extract samples based on a 1,000 µg/ml extract load.

The concentrations of CD200 were measured in triplicates, interpolated from the CD200 standard curve and corrected for sample dilution. Undiluted samples are SK-Mel-28 extract 20% and NCI-H226 extract 5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean CD200 concentration was determined to be 3,860 pg/ml in SK-Mel-28 extract and 21,265 pg/ml in NCI-H226 extract.

Interpolated concentrations of spiked CD200 in human cell culture media samples.

The concentrations of CD200 were measured in triplicates, interpolated from the CD200 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=3).