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[采用二辛可宁酸-Cu⁺比色反应的毛细管电泳法测定蛋白质]

[Capillary electrophoretic method for protein determination using bicinchoninic acid-Cu+ colorimetric reaction].

作者信息

Meng Qingwei, Guo Lei, Shen Rui, Xie Jianwei

机构信息

Beijing Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China.

出版信息

Se Pu. 2010 Jul;28(7):682-7. doi: 10.3724/sp.j.1123.2010.00682.

Abstract

A new rapid, sensitive method for protein determination using capillary electrophoresis and specific colorimetric reaction of bicinchoninic acid (BCA) was established, assisted by microwave incubation. With 60 mmol/L boric acid buffer (pH 9.5) and inclusion additive of beta-cyclodextrin, the complex of BCA-Cu+ and free BCA molecules were efficiently separated. The peak intensity of BCA-Cu+ was higher than those of native proteins about two orders of magnitude at a low wavelength of 200 nm. The linear ranges of this method were from 2 to 200 mg/L for transferrin, and 2 to 100 mg/L for ricin. The limits of detection for transferrin and ricin were 0.33 and 0.37 mg/L, respectively. This method was also successfully applied in the determination of some ricin samples in the First International Proficiency Test on the quantification of ricin.

摘要

建立了一种新的快速、灵敏的蛋白质测定方法,该方法采用毛细管电泳结合二辛可宁酸(BCA)特异性比色反应,并借助微波孵育。使用60 mmol/L硼酸缓冲液(pH 9.5)和β-环糊精包合添加剂,可有效分离BCA-Cu+络合物和游离BCA分子。在200 nm的低波长下,BCA-Cu+的峰强度比天然蛋白质高约两个数量级。该方法对转铁蛋白的线性范围为2至200 mg/L,对蓖麻毒素为2至100 mg/L。转铁蛋白和蓖麻毒素的检测限分别为0.33和0.37 mg/L。该方法还成功应用于第一次国际蓖麻毒素定量能力验证试验中一些蓖麻毒素样品的测定。

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