Biotin Conjugated Human CCL20/MIP-3 alpha Recombinant Rabbit Monoclonal Antibody [PSH12-33] - Detector

Sandwich ELISA analysis of Human CCL20/MIP-3 alpha matched pair antibodies

Capture: HA725010, Human CCL20/MIP-3 alpha Rabbit mAb [PSH12-32]
Detector: HA725011, Human CCL20/MIP-3 alpha Rabbit mAb [PSH12-33]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA725010) 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 CCL20/MIP-3 alpha protein (HA211012) starting from 2000 pg/ml to 0 pg/ml and detect antibody (HA725011, 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 CCL20 in THP-1 supernatant treated or untreated with Hu-IFN-r and LPS for overnight.

Capture: HA725010, Human CCL20/MIP-3 alpha Rabbit mAb [PSH12-32]
Detector: HA725011, Human CCL20/MIP-3 alpha Rabbit mAb [PSH12-33]

Interpolated concentration of native CCL20 was measured in duplicate at different sample concentrations and interpolated from the CCL20 standard curves. Undiluted samples were 50% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CCL20 concentration was determined to be 2,126 pg/mL in neat THP-1 treated supernatant, undetectable in untreated THP-1 supernatant.