Development of two dimensional Zymogram/SDS-PAGE separation technique in order to characterize and analyze degradation products of Type I Collagen by Trypsin and the Matrix Metalloproteinases known as gelatinases (MMP-2 and MMP-9) and Stromelysin (MMP-3)
B.S. (Bachelor of Science)
Department of Biological Sciences
Through manipulations of different variables and development of certain technical processes, it was possible to develop a two-dimensional separation technique for the degradation products of Type I Collagen. The first step was a zymogram method in which gelatin and vitrogen were degraded by both trypsin samples and matrix metalloproteinases. The zymogram gels were then run electrophoretically and then separated into various strips which were treated with 2.5% Triton X-100 and Buffer C and then allowed to incubate at various times and temperatures. The second step was an electrophoretic run of one of the excised strips from the original 2ymograms. Through the use of trypsin degradation and comparison of experimental data with deduced theoretical results, it was possible to validate the two-dimensional technique as viable. The two-dimensional technique with matrix metalloproteinases showed great promise as a characterization technique in that, with the optimal conditions, a good deal of detailed degradation product can be observed and analyzed for each individual matrix metalloproteinase.
Nigro, Kenneth J., "Development of two dimensional Zymogram/SDS-PAGE separation technique in order to characterize and analyze degradation products of Type I Collagen by Trypsin and the Matrix Metalloproteinases known as gelatinases (MMP-2 and MMP-9) and Stromelysin (MMP-3)" (1994). Honors Capstones. 403.
37 unnumbered pages
Northern Illinois University
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