Human CD62L Recombinant Rabbit Monoclonal Antibody [PSH13-86] - BSA and Azide free (Detector)

Sandwich ELISA analysis of human CD62L matched pair antibodies

Capture: HA723576, Human CD62L Rabbit mAb [PSH13-85]
Detector: HA723578, Human CD62L Rabbit mAb [PSH13-86]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723576) 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 CD62L protein (HA211181) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723578, 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 CD62L in human samples.

Capture: HA723576, Human CD62LRabbit mAb [PSH13-85]
Detector: HA723578, Human CD62L Rabbit mAb [PSH13-86]

Interpolated concentration of native CD62L was measured in duplicate at different sample concentrations and interpolated from the CD62L standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD62L concentration was determined to be 6,792 pg/mL in Jurkat cell supernatant. There was no detectable signal in Hela cell supernatant.

Interpolated concentrations of spiked CD62L in cell culture media samples.

Capture: HA723576, Human CD62L Rabbit mAb [PSH13-85]
Detector: HA723578, Human CD62L Rabbit mAb [PSH13-86]

The concentrations of CD62L were measured in duplicates, interpolated from the CD62L 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).