Human CCL3 / MIP-1 alpha Recombinant Rabbit Monoclonal Antibody [PSH11-76] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human CCL3 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723377) 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 CCL3 / MIP-1 alpha protein (HA210901) starting from 1000 pg/ml to 0 pg/ml and detect antibody (HA723378, 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 CLL3 in human THP-1 supernatant treated or untreated with PMA and LPS.

Interpolated concentration of native CLL3 was measured in duplicate at different sample concentrations and interpolated from the CLL3 standard curves. Undiluted samples are as follows: untreated 100% and treated 12.5%. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CLL3 concentration was determined to be 27 pg/ml in untreated THP-1 cell culture supernatant and 5,007 pg/mL in treated THP-1 supernatant.

Interpolated concentrations of spiked CLL3 in cell culture media samples.

The concentrations of CLL3 were measured in duplicates, interpolated from the CLL3 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).