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This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for MMP-2 and MMP-9 has been pre-coated onto a microplate. Standards and samples after 10-fold dilution, were pipetted (50?��l) into the wells to allow the link of specific molecules to immobilised antibody. After washing away any unbound substance, 200?��l of an enzyme-linked monoclonal antibody specific for MMP-2 or MMP-9 was added to the wells. After removing any unbound antibody-enzyme reagent, 200?��l of a substrate solution was added to the wells and colour developed in proportion to the amount of MMP2 and MMP9. The reaction was stopped with 50?��l HCl and the intensity Erlotinib in vitro of the colour was measured at 450?nm. The dilution factor was recorded to calculate the concentration of MMP2 and MMP9 in the samples. Data are expressed as mean?��?SD and analysed by Student's t-test. Differences were considered statistically significative at p??0.05). In particular, MMP-2 concentration Ponatinib was 127.15?��?45.56?SD?ng/ml, whilst MMP-9 concentration was 149.41?��?53.61?SD?pg/ml. In parallel with subacromial fluid levels, peripheral blood level of MMP-2 and Calpain MMP-9 were measured and the results are shown in Fig. 1, panels B and C. Panel B shows that peripheral blood levels of MM-2 (130.75?��?47.48?ng/ml) were comparable to that of the subacromial fluid (127.15?��?45.56?ng/ml), with no statistical significant difference (p?=?0.395). On the contrary, the concentration of MMP-9, as shown in panel C, was higher in the subacromial space (149.41?��?53.61?ng/ml) than in the peripheral blood (67.61?��?12.62?pg/ml). This difference was statistically significant, p value?