Analytical Sciences

Abstract − Analytical Sciences, 29(2), 255 (2013).

An Improved Protocol for Better Detection of Protein Using 8-Anilino-1-naphthalenesulfonate
Weide MA,*,** Guanghui ZHU,*** Ting ZHAO,* Weitao CONG,* Weijian YE,* and Litai JIN*
*Zhejiang Provincial Key Laboratory of Biopharmaceuticals, Wenzhou Medical College, Wenzhou 325035, P. R. China
**Laboratory of Gynecology and Obstetrics, People’s Hospital of Wenzhou, Wenzhou 325035, P. R. China
***The Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325035, P. R. China
This study describes the optimum conditions at the staining time and the signal intensity for using 8-anilino-1-naphthalenesulfonate (ANS) as a fluorescent probe to detect proteins in SDS-PAGE. Using the optimized protocol, protein can be easily detected by short time fixing (20 min) and washing (2 × 5 min), followed by 10 min of staining. As low as 1 – 2 ng of the protein band can be detected, approximately thirty-fold higher than that of the original protocol. Furthermore, the compatibility of the staining method with MS was also explored by comparing the peptide mass fingerprinting results data of serial dilutions of BSA and ovalbumin stained by ANS with SYPRO Ruby.