Human CD33 Recombinant Rabbit Monoclonal Antibody [PSH05-26] - BSA and Azide free (Detector)

Sandwich ELISA analysis of Human CD33 matched pair antibodie

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA722254) 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 CD33 protein (HA210949) starting from 10000 pg/ml to 0 pg/ml and detect antibody (HA722255, 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.

The concentrations of CD33 were interpolated from the CD33 standard curves and corrected for sample dilution. Undiluted samples are mixed serum from twenty-one volunteers 5%. The mean CD33 concentration was determined to be 7,466 pg/mL in mixed serum from twenty-one volunteers.

Interpolated concentrations of native CD33 in U937 and HL-60 extract samples based on a 1,000 µg/mL extract load.
The concentrations of CD33 were measured in triplicates, interpolated from the CD33 standard curve and corrected for sample dilution. Undiluted samples are U937 extract 5% and HL-60 extract 1%. The mean CD33 concentration was determined to be 5,333 pg/mL in U937 extract and 21,225 pg/mL in HL-60 extract.

Interpolated concentrations of spiked CD33 in human cell culture media samples.
The concentrations of CD33 were measured in triplicates, interpolated from the CD33 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).