Biotin Conjugated Human CD40 Recombinant Rabbit Monoclonal Antibody [PSH22-80] - Detector

Sandwich ELISA analysis of Human CD40 matched pair antibodies

Capture: HA724324, Human CD40 Rabbit mAb [PSH22-79]
Detector: HA724325, Human CD40 Rabbit mAb [PSH22-80]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724324) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 CD40 protein (HA211232) starting from 1000 pg/ml to 0 pg/ml and detect antibody (HA724325, 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 CD14 in Human serum.

Capture: HA724324, Human CD40 Rabbit mAb [PSH22-79]
Detector: HA724325, Human CD40 Rabbit mAb [PSH22-80]

Interpolated concentration of native CD14 was measured in duplicate at different sample concentrations and interpolated from the CD14 standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD14 concentration was determined to be 808 pg/ml in Human serum.

Interpolated concentrations of native CD40 in human samples.

Capture: HA724324, Human CD40 Rabbit mAb [PSH22-79]
Detector: HA724325, Human CD40 Rabbit mAb [PSH22-80]

Interpolated concentration of native CD40 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD40 concentration was determined to be 1,566 pg/mL in human urine and 41 pg/ml in Ramos cell culture supernatant.There was no detectable signal in Jurkat cell culture supernatant.

Interpolated concentrations of spiked CD40 in cell culture media samples.

Capture: HA724324, Human CD40 Rabbit mAb [PSH22-79]
Detector: HA724325, Human CD40 Rabbit mAb [PSH22-80]

The concentrations of CD40 were measured in duplicates, interpolated from the CD40 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).