Methods in Molecular Biology

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Volume 1 in this book describes how to determine the activity of different isozymes, allozymes, and families of proteinases to advance the fields of enzymology and molecular evolution, and provides useful biomarkers for various biological processes, pathological conditions, and clinical disorders. The chapters in Volume 1 are organized in four parts. Part I introduces gelatin zymography principle, method and precautions, and a practical view of zymography.  Part II provides details on the use of zymography to assess various endopeptidases, and includes updates on protease zymography, protocols for gelatin zymography, detection of gelatinases by substrate zymography, unveiling matrix metalloproteinase (MMP) dynamics, and measuring MMPs activity by gelatin zymography. Part III includes chapters on one- and two-dimensional zymography for assessing proteases, loading-controlled zymography, and the use of gel diffusion-based gelatin zymography to analyze MMP-2 and MMP-9 activity in cell culture. Part IV focuses on the use of zymography to assess protease inhibitors and includes chapters that describe polyacrylamide gel electrophoresis as a tool to identify potential inhibitors of protein glycation and analyze glycation-induced protein cross-linking inhibition, kinetics of protease thermal inactivation, and reversible inhibition of cysteine proteases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and thorough, Zymography: Principles, Process, and Pitfalls, Volume I is a valuable resource for both experts in the field, as well as new scientists aspiring to learn and perform successful zymography techniques.