Human CD137 Recombinant Rabbit Monoclonal Antibody [PSH08-63] - BSA and Azide free (Capture)

Sandwich ELISA analysis of Human CD137 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723012) 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 CD137 protein (HA210839) starting from 8,000 pg/ml to 0 pg/ml and detect antibody (HA723013, 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 CD137 in HDLM-2, Jurkat and MOLT-4 extract samples based on a 2,000 µg/ml extract load.

The concentrations of CD137 were measured in duplicates, interpolated from the CD137 standard curve and corrected for sample dilution. Undiluted samples are HDLM-2 extract 100%, Jurkat extract 100% and HOLT-4 extract 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD137 concentration was determined to be 269.6 pg/ml in HDLM-2 extract and undetectable in Jurkat and MOLT-4 extract.

Interpolated concentrations of spiked CD137 in human cell culture media samples.

The concentrations of CD137 were measured in duplicates, interpolated from the CD137 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).