COLLAGEN ZYMOGRAPHY AS A SENSITIVE AND SPECIFIC TECHNIQUE FOR THE DETERMINATION OF SUBPICOGRAM LEVELS OF INTERSTITIAL COLLAGENASE

We describe the use of casting native collagen type I in SDS-polyacryl amide gel (collagen zymography) for the determination of interstitial collagenase. As with gelatin, the incorporation of collagen in the gel s reduced protein migration and the need for making corrections for an accurate M-r evaluation. This method proved to be very sensitive: 0.1 pg of APMA-activated procollagenase could be detected, and specific l evels of active gelatinase or stromelysin lower than 5 ng were inactiv e under our experimental conditions. It was used to demonstrate the in creased expression of collagenase following treatment of human gingiva l fibroblasts with interleukin-1 beta; the amounts of enzyme quantifie d by either collagen zymography or immunodot blot assay are comparable . (C) 1998 Academic Press.