通过考马斯亮蓝和固绿的红外荧光对凝胶和印迹上的蛋白质进行定量分析。

Quantitation of protein on gels and blots by infrared fluorescence of Coomassie blue and Fast Green.

作者信息

Luo Shen, Wehr Nancy B, Levine Rodney L

机构信息

Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, Bethesda, MD 20892, USA.

出版信息

Anal Biochem. 2006 Mar 15;350(2):233-8. doi: 10.1016/j.ab.2005.10.048. Epub 2005 Nov 17.

DOI:10.1016/j.ab.2005.10.048

PMID:16336940

Abstract

Coomassie blue staining of gels and blots is commonly employed for detection and quantitation of proteins by densitometry. We found that Coomassie blue or Fast Green FCF bound to protein fluoresces in the near infrared. We took advantage of this property to develop a rapid and sensitive method for detection and quantitation of proteins in gels and on blots. The fluorescence response is quantitative for protein content between 10 ng and 20 microg per band or spot. Staining and destaining require only 30 min, and the method is compatible with subsequent immunodetection.

摘要

凝胶和印迹的考马斯亮蓝染色通常用于通过光密度测定法检测和定量蛋白质。我们发现与蛋白质结合的考马斯亮蓝或固绿FCF在近红外区域发出荧光。我们利用这一特性开发了一种快速灵敏的方法，用于检测和定量凝胶和印迹中的蛋白质。荧光响应对于每条带或每个斑点中10 ng至20 μg的蛋白质含量是定量的。染色和脱色仅需30分钟，并且该方法与后续的免疫检测兼容。