You W W, Haugland R P, Ryan D K, Haugland R P
Molecular Probes, Inc., Eugene, Oregon 97402, USA.
Anal Biochem. 1997 Jan 15;244(2):277-82. doi: 10.1006/abio.1996.9920.
This paper describes the application of 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA), a sensitive fluorogenic reagent for detection of amines, to the assay of proteins in solution. The sensitivity and dynamic range of CBQCA in the determination of protein concentration is modulated by the number of accessible amines present in the protein assayed. Thus, this method bears the same limitations as other reagents used in fluorogenic assays based on fluorescent amine adducts such as those obtained with fluorescamine or o-phthaldialdehyde, or of other spectrophotometric methods, where the color development is determined by the abundance in the protein of certain amino acids. However, in comparison to other fluorescence-based protein detection methods, CBQCA has proven to be an extremely sensitive reagent with a very broad, essentially linear dynamic range, capable of detecting from 10 ng to 150 micrograms of protein (in a 100- to 200-microL assay volume). The CBQCA reagent also functions well in the presence of substances, such as lipids, known to interfere in many other protein determination methods.
本文描述了3-(4-羧基苯甲酰基)喹啉-2-甲醛(CBQCA)，一种用于检测胺类的灵敏荧光试剂，在溶液中蛋白质测定中的应用。CBQCA在蛋白质浓度测定中的灵敏度和动态范围受被测蛋白质中可及胺数量的调节。因此，该方法与基于荧光胺加合物的荧光测定中使用的其他试剂，如与荧光胺或邻苯二甲醛获得的试剂，或其他分光光度法具有相同的局限性，在这些方法中，显色由蛋白质中某些氨基酸的丰度决定。然而，与其他基于荧光的蛋白质检测方法相比，CBQCA已被证明是一种极其灵敏的试剂，具有非常宽的、基本呈线性的动态范围，能够检测10 ng至150 μg的蛋白质(在100至200 μL的测定体积中)。CBQCA试剂在存在已知会干扰许多其他蛋白质测定方法的物质(如脂质)的情况下也能很好地发挥作用。
