Carvalho-Silva Jônatas, Fernandes Lígia M G, Converti Attilio, Porto Tatiana S
Departament of Morphology and Animal Physiology, Federal Rural University of Pernambuco, Recife, Pernambuco, Brazil.
Department of Civil, Chemical and Environmental Engineering, University of Genova, Genoa, Italy.
Methods Mol Biol. 2025;2917:109-120. doi: 10.1007/978-1-0716-4478-2_10.
Zymography is an electrophoretic technique used to demonstrate enzyme activity during separation in polyacrylamide gels under nonreducing conditions. This kind of analysis can be used in association with electrophoresis (SDS-PAGE) to show which protein is correlated with a given activity. Proteases are the most used enzymes for this technique since the polyacrylamide gel can be impregnated with azocasein, a casein derivative with an added sulfanilamide group (orange color). Furthermore, azocasein can be degraded while the protease is separated through the gel. The degradation is easily revealed by dyeing the azocasein with Coomassie blue, with the nonblue area in the gel being the zone where the separated protease shows activity. Zymography has been widely used for the identification and characterization of new proteolytic enzymes, but it can detect latent and active forms of enzymes in cells, tissues, or biological fluids.
酶谱法是一种电泳技术，用于在非还原条件下于聚丙烯酰胺凝胶中分离过程中展示酶活性。这种分析可与电泳（SDS-PAGE）结合使用，以显示哪种蛋白质与特定活性相关。蛋白酶是该技术最常用的酶，因为聚丙烯酰胺凝胶可浸渍偶氮酪蛋白，一种添加了磺胺基团（橙色）的酪蛋白衍生物。此外，在蛋白酶通过凝胶分离时，偶氮酪蛋白会被降解。通过用考马斯亮蓝对偶氮酪蛋白染色，很容易揭示这种降解，凝胶中未变蓝的区域就是分离出的蛋白酶显示活性的区域。酶谱法已广泛用于新蛋白水解酶的鉴定和表征，但它可以检测细胞、组织或生物体液中酶的潜在形式和活性形式。
