Biotin Conjugated Human CXCL5 Recombinant Rabbit Monoclonal Antibody [PSH18-71] - Detector

Sandwich ELISA analysis of human CXCL5 matched pair antibodies

Capture: HA725361, Human CXCL5 Rabbit mAb [PSH18-70]
Detector: HA725362, Human CXCL5 Rabbit mAb [PSH18-71]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725361) diluted in carbonate/bicarbonate buffer, at a concentration of 2 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 CXCL5 protein (HA210887) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA725362, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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 37℃ in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Interpolated concentrations of native CXCL5 in HeLa cell culture supernatant and human plasma samples.

Capture: HA725361, Human CXCL5 Rabbit mAb [PSH18-70]
Detector: HA725362, Human CXCL5 Rabbit mAb [PSH18-71]

The concentrations of CXCL5 were measured in duplicates, interpolated from the CXCL5 standard curve and corrected for sample dilution. Undiluted samples are HEK-293 cell culture supernatant 100% and human plasma 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CXCL5 concentration was determined to be 6375 pg/ml in human plasma and undetectable in HEK-293 cell culture supernatant.