Biotin Conjugated Human CCL17 / TARC Recombinant Rabbit Monoclonal Antibody [PSH13-34] - Detector

Sandwich ELISA analysis of Human CCL17 / TARC matched pair antibodies

Capture: HA723513, Human CCL17 / TARC Rabbit mAb [PSH13-33]
Detector: HA723514, Human CCL17 / TARC Rabbit mAb [PSH13-34]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723513) 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 Human CCL17 / TARC protein (HA210895) starting from 2,000 pg/ml to 0 pg/ml and detect antibody (HA723514, 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 CCL17 in human PBMC cell culture supernatant.

Capture: HA723513, Human CCL17 / TARC Rabbit mAb [PSH13-33]
Detector: HA723514, Human CCL17 / TARC Rabbit mAb [PSH13-34]

PBMC cells were stimulated with 10 µg/ml PHA-M and incubated for 5 days. The concentrations of CCL17 measured in duplicate and interpolated from the CCL17 standard curve and corrected for sample dilution. Undiluted samples are as follows: stimulated 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CCL17 concentration was determined to be 8,995 pg/ml in PHA-M stimulated PBMC cell culture supernatant.

Interpolated concentrations of spiked CCL17 in human cell culture media samples.

Capture: HA723513, Human CCL17 / TARC Rabbit mAb [PSH13-33]
Detector: HA723514, Human CCL17 / TARC Rabbit mAb [PSH13-34]

The concentrations of CCL17 were measured in duplicates, interpolated from the CCL17 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).