Nešporová Kristina, Lehká Kateřina, Musilová Pavlína
Contipro a.s., Dolní Dobrouč 401, Dolní Dobrouč, Czech Republic.
Faculty of Chemical Technology, University of Pardubice, Pardubice 2, Czech Republic.
Methods Mol Biol. 2025;2917:143-150. doi: 10.1007/978-1-0716-4478-2_13.
Hyaluronan and hyaluronidases are critical in tissue remodeling, inflammation, and tumor progression. This chapter provides a comprehensive guide to hyaluronan zymography, a powerful technique for detecting and quantifying hyaluronidase activity in complex biological samples. The method involves separating proteins by polyacrylamide gel electrophoresis with hyaluronan incorporated into the gel matrix. Following electrophoresis, the gel is incubated to allow hyaluronidases to degrade the hyaluronan substrate, resulting in clear digestion zones. Detailed protocols for sample preparation and the zymographic process are included, offering researchers a robust tool for studying hyaluronidase activity and regulation in various biological contexts.
透明质酸和透明质酸酶在组织重塑、炎症和肿瘤进展中起着关键作用。本章提供了一份关于透明质酸酶谱法的全面指南，这是一种用于检测和定量复杂生物样品中透明质酸酶活性的强大技术。该方法包括通过聚丙烯酰胺凝胶电泳分离蛋白质，并将透明质酸掺入凝胶基质中。电泳后，将凝胶孵育以使透明质酸酶降解透明质酸底物，从而产生清晰的消化区。文中包含了样品制备和酶谱分析过程的详细方案，为研究人员提供了一个强大的工具，用于研究透明质酸酶在各种生物学环境中的活性和调控。
