Biotin Conjugated Human SCF Recombinant Rabbit Monoclonal Antibody [PSH07-58]

Sandwich ELISA analysis of human SCF matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722871) 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 SCF protein (HA210618) starting from 2000 pg/ml to 0 pg/ml and detect antibody (HA722873B, 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 SCF in human cell culture supernatant samples.

Interpolated concentration of native SCF was measured in duplicate at different sample concentrations and interpolated from the SCF standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean SCF concentration was determined to be 759 pg/mL in HT-29 supernatant, undetectable in K-562 in supernatant.

Interpolated concentrations of spiked SCF in cell culture media samples.

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