Biotin Conjugated Human CCL21 Recombinant Rabbit Monoclonal Antibody [PSH13-37] - Detector

Sandwich ELISA analysis of human CCL21 matched pair antibodies

Capture: HA723518, Human CCL21 Rabbit mAb [PSH13-36]
Detector: HA723519, Human CCL21 Rabbit mAb [PSH13-37]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723518) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 CCL21 protein (HA211144) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA723519, 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 CCL21 in human cell culture supernatant samples.

Capture: HA723518, Human CCL21 Rabbit mAb [PSH13-36]
Detector: HA723519, Human CCL21 Rabbit mAb [PSH13-37]

Interpolated concentration of native CCL21 was measured in duplicate at different sample concentrations. Undiluted samples were 50% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CCL21 concentration was determined to be 1,336 pg/mL in human serum.

Interpolated concentrations of spiked CCL21 in cell culture media samples.

Capture: HA723518, Human CCL21 Rabbit mAb [PSH13-36]
Detector: HA723519, Human CCL21 Rabbit mAb [PSH13-37]

The concentrations of CCL21 were measured in duplicates, interpolated from the CCL21 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).