Human M-CSF Recombinant Rabbit Monoclonal Antibody [PSH11-37] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human M-CSF matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723324) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 M-CSF protein (HA210918) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA72332, 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 M-CSF in MG-63 cell culture supernatant and human urine samples.

The concentrations of M-CSF were measured in duplicates, interpolated from the M-CSF standard curve and corrected for sample dilution. Undiluted samples are MG-63 cell culture supernatant 20% and human urine 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean M-CSF concentration was determined to be 16,640 pg/ml in MG-63 cell culture supernatant and 738 pg/ml in human urine samples.

Interpolated concentrations of spiked M-CSF in human cell culture media samples.

The concentrations of M-CSF were measured in duplicates, interpolated from the M-CSF 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).