Li X, Abdi K, Mentzer S J
Dana-Farber Cancer Institute, Brigham and Women's Hospital, Boston, MA 02115.
J Immunol Methods. 1994 Jun 24;172(2):141-5. doi: 10.1016/0022-1759(94)90101-5.
o-Phthaldehyde (OPA) reagent reacts with primary amines in aqueous solution at room temperature. When the reaction occurs in the presence of 2-mercaptoethanol, a bright blue fluorescence is produced. We investigated the application of the OPA reagent in the routine determination of monoclonal antibody concentration. The OPA reagent produced maximal fluorescence within 30 min and was stable for 2 h. Buffers containing primary amines gave high background fluorescence; however, the OPA reagent gave reliable results with the most commonly used antibody solutions. The OPA reagent provided a rapid and simple measure of protein concentration from 1 microgram to 1 mg and was readily adapted for use with a 96-well fluorescence reader.
邻苯二甲醛（OPA）试剂在室温下能与水溶液中的伯胺发生反应。当反应在2-巯基乙醇存在的情况下发生时，会产生亮蓝色荧光。我们研究了OPA试剂在单克隆抗体浓度常规测定中的应用。OPA试剂在30分钟内产生最大荧光，并且在2小时内保持稳定。含有伯胺的缓冲液会产生高背景荧光；然而，OPA试剂与最常用的抗体溶液反应时能给出可靠的结果。OPA试剂能快速简便地测定1微克至1毫克的蛋白质浓度，并且很容易适配96孔荧光读数仪使用。
