Biotin Conjugated Human CHIT1 Recombinant Rabbit Monoclonal Antibody [PSH15-90] - Detector

Sandwich ELISA analysis of human CHIT1 matched pair antibodies

Capture: HA725205, Human CHIT1 Rabbit mAb [PSH15-89]
Detector: HA725206, Human CHIT1 Rabbit mAb [PSH15-90]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA725205) 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 CHIT1 protein (HA211215) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA725206, 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 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 CHIT1 in HeLa cell culture supernatant and human plasma.

Capture: HA725205, Human CHIT1 Rabbit mAb [PSH15-89]
Detector: HA725206, Human CHIT1 Rabbit mAb [PSH15-90]

The concentrations of CHIT1 were measured in triplicates, interpolated from the CHIT1 standard curve and corrected for sample dilution. Undiluted samples are HeLa cell culture supernatant 50% and human plasma 2.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean CHIT1 concentration was determined to be 13787 pg/ml in human plasma and undetectable in HeLa cell culture supernatant.