Phorbol Ester Induced Upregulation of MMP-3 and MMP-9 in HT1080 Cells. Levels of MMP-3 (using Cat. No. QIA73) and MMP-9 (using Cat. No. QIA56) were measured following treatment with phorbol 12-myristate 13-acetate (PMA) as outlined in the respective Detailed Protocol.

Phorbol Ester Induced Upregulation of MMP-3 in MG-63 Cells. Levels of MMP-3 were measured in MG-63 cells following treatment with PMA for 96 h as outlined in the Detailed Protocol.

Sensitivity. The lower limit of detection was measured by assaying three replicates of zero four times using two different lots of plates and two different lots of detector antibody. The grand mean signal and pooled standard deviation of zero was calculated. The grand mean of each standard (run in replicates of three in the four assays) was used for the standard curve, and the response, mean signal of zero plus two standard deviations, read in dose from the standard curve is the LLD; that is, the smallest dose that is not zero with 95% confidence.

Specificity. Levels of MMP-3 in human serum (top) and cell culture supernatants of CCF-STTG1 astrocytoma cells (bottom) were measured following immunaffinity extraction with three MMP-3 antibodies (that are not used in this ELISA kit) and TIMP antibodies. PBS was used as the no-antibody control.

Specificity. The levels of human MMP-3/TIMP complex (8 ng/ml; purified protein) and MMP-3 (8 ng/ml; purified protein) were measured as outlined in the Detailed Protocol.