Cheung A L, Ying P, Fischetti V A
Laboratory of Bacteriology and Immunology, Rockefeller University, New York, New York 10021.
Anal Biochem. 1991 Feb 15;193(1):20-3. doi: 10.1016/0003-2697(91)90037-t.
Routine assays to detect proteinases in biological samples are generally tedious and time-consuming. To expedite the recognition of proteinases, we have developed an assay utilizing the gelatin on the surface of an unprocessed Kodak X-Omat AR film as the proteolytic substrate. A positive reaction is indicated by a clear zone on the film after it has been rinsed with running water. This proteinase assay has been found to be inexpensive, rapid, and simple. Besides its ease of use, this assay has been found to be quantitatively reproducible with a well-defined endpoint. More importantly, this assay method is applicable to a variety of proteolytic enzymes under diverse pH (5-8.5) and salt conditions (up to 5 M NaCl) and has a sensitivity similar to that of azocoll. Since the assay does not require sophisticated equipment, it is useful as a general laboratory procedure.
检测生物样品中蛋白酶的常规分析方法通常既繁琐又耗时。为了加快蛋白酶的识别，我们开发了一种分析方法，利用未处理的柯达X-Omat AR胶片表面的明胶作为蛋白水解底物。用流水冲洗胶片后，胶片上出现的透明区域表明反应呈阳性。已发现这种蛋白酶分析方法价格低廉、快速且简单。除了使用方便外，还发现该分析方法在明确的终点下具有定量可重复性。更重要的是，该分析方法适用于多种蛋白酶，适用于不同的pH值（5 - 8.5）和盐浓度条件（高达5 M氯化钠），并且灵敏度与偶氮酪蛋白类似。由于该分析方法不需要复杂的设备，因此作为一种通用的实验室程序很有用。
