Human CXCL1/GRO alpha Recombinant Rabbit Monoclonal Antibody [PSH05-94] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human CXCL1 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722493) 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 standard Human CXCL1/GRO alpha protein (HA210838) starting from 2000 pg/ml to 0 pg/ml and detect antibody (HA722494, 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 CXCL1 in human PBMC culture supernatant samples.

Human PBMCs were cultured for 5 days at 37˚C in the presence or absence of 10ug/ml PHA-M. The concentrations of CXCL1 were interpolated from the standard curve and corrected for sample dilution. The mean CXCL1 concentration was determined to be 32pg/mL in unstimulated PBMC supernatants, and 1745 pg/mL in stimulated PBMC supernatants.

Interpolated concentrations of spiked CXCL1 in human cell culture media samples.

The concentrations of CXCL1 were measured in duplicates, interpolated from the CXCL1 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).