Micocci Kelli C, Stotzer Uliana S, Moritz Milene N O
Department of Chemistry, Federal University of São Carlos, Sao Paulo, Brazil.
Human Movement Sciences Research Group, Methodist University of Piracicaba, Sao Paulo, Brazil.
Methods Mol Biol. 2025;2917:65-74. doi: 10.1007/978-1-0716-4478-2_6.
Gelatin zymography is a widely popular method due to its simplicity, low cost, and quick results, for studying gelatinases in various biological systems. Zymography can detect both the pro and active forms of matrix metalloproteinases MMP-2 (gelatinase A) and MMP-9 (gelatinase B). These MMPs play critical roles in the pathophysiology of many human diseases, particularly in cancer progression. Gelatin zymography is a method based on a suitable protein substrate incorporated into a sodium dodecyl sulfate-polyacrylamide gel. Substrate degradation by protease-containing samples can be visualized through the contrast between the Coomassie blue-stained gel and the white band of substrate degradation. Here, we provide a straightforward, step-by-step methodology for detecting MMP-2 and MMP-9 gelatinases in tumor cells. It is essential to highlight that accurately interpreting the data requires a thorough understanding of the technique's principles.
明胶酶谱法因其操作简单、成本低且结果快速，成为研究各种生物系统中明胶酶的一种广泛流行的方法。酶谱法可以检测基质金属蛋白酶MMP-2（明胶酶A）和MMP-9（明胶酶B）的前体形式和活性形式。这些基质金属蛋白酶在许多人类疾病的病理生理学中起着关键作用，尤其是在癌症进展过程中。明胶酶谱法是一种基于将合适的蛋白质底物掺入十二烷基硫酸钠-聚丙烯酰胺凝胶中的方法。含蛋白酶的样品对底物的降解可以通过考马斯亮蓝染色凝胶与底物降解的白色条带之间的对比来可视化。在这里，我们提供了一种直接的、分步的方法来检测肿瘤细胞中的MMP-2和MMP-9明胶酶。必须强调的是，准确解读数据需要对该技术的原理有透彻的理解。
