Human MMP-8 Recombinant Rabbit Monoclonal Antibody [PSH03-33] - BSA and Azide free (Capture)

Sandwich ELISA analysis of human MMP8 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA721978) 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 human MMP8 protein (HA210646) starting from 5000 pg/ml to 0 pg/ml and detect antibody (HA721979, 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.

The concentrations of MMP8 were measured in duplicates, interpolated from the MMP8 standard curves and corrected for sample dilution. The mean MMP8 concentration was determined to be 1043.74 pg/mL in A375 supernatant.

The concentrations of MMP8 were measured in duplicates, interpolated from the MMP8 standard curves and corrected for sample dilution. The mean MMP8 concentration was determined to be 24841.38 pg/mL in SK-Mel-28 supernatant.

The concentrations of MMP8 were measured in duplicates, interpolated from the MMP8 standard curves and corrected for sample dilution. Undiluted sample is mixed human serum from twenty volunteers 1%. The mean MMP8 concentration was determined to be 240.35 ng/mL in mixed human serum from twenty volunteers.