MMP 2/9 zymographic analysis in breast cancer and surrounding tissues.
The cytosols fromtumorand its corresponding surrounding tissue were assayed for MMP activity. One percent type B-gelatin solution prepared in water, 8 % resolving gel is prepared containing 1% B-gelatin solution. Samples were prepared in MMP dye and loaded in gel to run at 110V for 60 minute. After completion of electrophoresis the gel wasactivated by washing buffer (as mentioned in the referenced protocol) and then incubated in zymographic gel development buffer pH 7.4 at 37ÂșC for 42 hours. Finally the gel was stained in amide black or Coommasie blue solution for 1 hour at room temperature in a shaker. The gels were then destained with solution I for 15 to 30 minutes, followed by destaining solution II for 3 to 5 hours until clear MMP bands appeared. After washing the destaining solution the gel was incubated in a gel preservative solution for 15 minutes and finally scanned with a digital scanner (21)