Volume 2 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 2 are organized in three parts. Part I introduces in situ zymography and localization of bright green-fluorescent gelatinase activity in tissue sections, in situ zymography in formalin-fixed paraffin-embedded and mineralized tissues, and in vivo zymography as an essential activity assay for studying the activity of matrix metalloproteinases (MMPs) in a cell-specific manner in the brain. Part II focuses on biological applications of zymography such as fundamentals of zymography and its applications to the study of biological samples, gelatin zymography to quantify MMP-2 and MMP-9 in complex biological specimens, and detection of proteolytic enzymes in polyacrylamide gels supplemented with diverse biological substrates. Part III focuses on potential clinical applications of zymography, with chapters describing assessment of MMP-2 and MMP-9 hydrolytic activity in preclinical and clinical tissue samples, the use of zymography to assess circulating MMP-2 and MMP-9 in plasma and serum and in pathological conditions, and the use of zymography for the detection of bacterial 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 prools, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Zymography: Biological and Clinical Applications, Volume II is a valuable resource for both experts in the field, as well as new scientists aspiring to learn and perform successful zymography techniques.