Validation of gelatin zymography.

<p><b>A.</b> Dilution series of MMP-2 standard. A standard curve was generated from human full length MMP-2 from 0–2.5 ng of total MMP-2. Pixel density measured from zymograms showed a linear relationship with the MMP-2 amount. <b>B.</b> Representative zymogram of standard dilution series reveals decreasing signal strength in conjunction with declining MMP-2 amount. <b>C.</b> Dilution series of protein extracts gained from patients with ascending aortic aneurysms. Protein extracts were diluted from 2fold until 16fold. The measured pixel density in the zymograms showed a linear decrease at dilution factors between 2fold and 8fold. Measurements of 4 independent dilutions illustrated the assay`s reproducibility. Each value represents the mean of two samples at the same dilution measured in the same gel. <b>D.</b> Representative zymogram of protein extract dilution series shows decreasing signal strength in conjunction with an increasing dilution factor. <b>E.</b> Representative serum and tissue zymograms showing different gelatinolytic activities. serum zymogram shows gelatinolytic activities corresponding to pro-MMP-2. Tissue zymogram shows gelatinolytic activities corresponding to pro-MMP2 and active MMP-2. Westernblot confirmed that the detected bands were MMP-2. <b>1,2</b>: patient sample. <b>S:</b> Human full length MMP-2.