Biotin Conjugated Mouse CD86 Recombinant Rabbit Monoclonal Antibody [PSH13-64] - Detector

Sandwich ELISA analysis of Mouse CD86 matched pair antibodies

Capture: HA723552, Mouse CD86 Rabbit mAb [PSH13-63]
Detector: HA723553, Mouse CD86 Rabbit mAb [PSH13-64]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723552) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 Mouse CD86 protein (HA210879) starting from 1,000 pg/ml to 0 pg/ml and detect antibody (HA723553, 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 CD86 in mouse serum samples.

Capture: HA723552, Mouse CD86 Rabbit mAb [PSH13-63]
Detector: HA723553, Mouse CD86 Rabbit mAb [PSH13-64]

Interpolated concentration of native CD86 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD86 concentration was determined to be 999 pg/mL in Mouse serum.

Interpolated concentrations of native CD68 in J774A.1 Cell extract samples based on a 1000 µg/ml extract load.

Capture: HA723552, Mouse CD86 Rabbit mAb [PSH13-63]
Detector: HA723553, Mouse CD86 Rabbit mAb [PSH13-64]

Interpolated concentration of native CD86 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD86 concentration was determined to be 220 pg/mL in J774A.1 Cell extract.

Interpolated concentrations of spiked CD86 in cell culture media samples.

Capture: HA723552, Mouse CD86 Rabbit mAb [PSH13-63]
Detector: HA723553, Mouse CD86 Rabbit mAb [PSH13-64]

The concentrations of CD86 were measured in duplicates, interpolated from the CD86 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).