Institute Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology, Jeonbuk, Korea.
Biotechnol Lett. 2011 Aug;33(8):1663-6. doi: 10.1007/s10529-011-0619-3. Epub 2011 Apr 13.
A new zymogram method, silver-stained fibrin zymography, for separation of protease bands and activity detection using a single substrate gel, was developed. The method takes advantage of the nanoscale sensitivity of both zymography and silver staining. After SDS-PAGE in a gel containing fibrin, the gel was incubated in enzyme reaction buffer and the zymogram was silver-stained. Bands with protease activity were stained with silver in clear areas where the protein substrate had been degraded. The molecular sizes of proteases were accurately determined. Furthermore, proteases of high molecular weight were clearly and sharply resolved.
开发了一种新的酶谱方法，即银染纤维蛋白酶谱法，用于在单一底物凝胶上分离蛋白酶带和活性检测。该方法利用了酶谱和银染的纳米级灵敏度。在含有纤维蛋白的凝胶中进行 SDS-PAGE 后，将凝胶在酶反应缓冲液中孵育，并用银染进行酶谱染色。在蛋白质底物被降解的透明区域，具有蛋白酶活性的条带被银染染色。准确测定了蛋白酶的分子量。此外，高分子量的蛋白酶也得到了清晰而锐利的分辨。
