Human MMP-3 Recombinant Rabbit Monoclonal Antibody [PSH07-62] - BSA and Azide free (Detector)

Sandwich ELISA analysis of human MMP3 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722877) 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 MMP-3 protein (HA210853) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA722878, 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 MMP3 in human serum samples.

The concentrations of MMP3 were interpolated from the MMP3 standard curves and corrected for sample dilution. Undiluted samples are human serum 20%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MMP3 concentration was determined to be 12,706 pg/ml in human serum.

Interpolated concentrations of native MMP3 in U-2 OS cell culture supernatant.

The concentrations of MMP3 were interpolated from the MMP3 standard curves and corrected for sample dilution. Undiluted samples are U-2 OS cell culture supernatant 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean MMP3 concentration was determined to be 4,120 pg/ml in U-2 OS cell culture supernatant.

Interpolated concentrations of spiked MMP3 in human cell culture media samples.

The concentrations of MMP3 were measured in duplicates, interpolated from the MMP3 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).