Fluorescent staining of protein in sodium dodecyl sulfate polyacrylamide gels by salicylaldehyde azine

• Published in: Electrophoresis Vol. 34; no. 22-23; pp. 3171 - 3179
• Main Authors: Ni, Mao-Wei, Ye, Wei-Jian, Cong, Wei-Tao, Hong, Guo-Ying, Zhu, Zhong-Xin, Duan, Yuan-Meng, Zhou, Xuan, Jin, Li-Tai
• Format: Journal Article
• Language: English
• Published: Germany 01.12.2013
• Subjects: Acrylic Resins - chemistry; Aldehydes - chemistry; Animals; Brain Chemistry; Cattle; Electrophoresis, Polyacrylamide Gel - methods; Fluorescent Dyes - chemistry; Mice; Molecular Docking Simulation; Proteins - analysis; Proteins - isolation & purification; Serum Albumin, Bovine - chemistry; Sodium Dodecyl Sulfate - chemistry
• ISSN: 1522-2683
• DOI: 10.1002/elps.201300241

As a non-covalent fluorescence probe, in this study, salicylaldehyde azine (SA) was introduced as a sensitive fluorescence-based dye for detecting proteins both in 1-D and 2-D polyacrylamide electrophoresis gels. Down to 0.2 ng of single protein band could be detected within 1 h, which similars to that of glutaraldehyde (GA)-silver stain, but approximately four times higher than that of SYPRO Ruby fluorescent stain. Furthermore, comparative analysis of the MS compatibility of SA stain with SYPRO Ruby stain indicated that SA stain is compatible with the downstream of protein identification by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Additionally, the probable mechanism of the SA stain was investigated by molecular docking. The results demonstrated that the interaction between SA and protein was mainly contributed by hydrogen bonding and hydrophobic forces.