Department of Biomedical Engineering, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, 518020 Guangdong, China.
Biomed Res Int. 2022 May 13;2022:6243971. doi: 10.1155/2022/6243971. eCollection 2022.
Silver staining is an excellent technique for detecting proteins that are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Protein silver staining technology has higher sensitivity and is suitable for the detection of low-concentration proteins compared to other staining techniques including the Coomassie brilliant blue detection method. The present study was conducted to enhance the detection ability of the protein staining method. Herein, we modified the recipe of silver staining, a very reproducible method, by adding AMP, PVP, Tween-80, and xylene to enhance the detection ability of protein staining. Furthermore, the particle size and potentiometer were used to detect the particle size and potential difference of the silver ions in the prepared dyeing materials, and then, the morphology, transparency, and size of the dyed silver particles in different dyeing solutions were studied using a transmission electron microscopy (TEM). The obtained results revealed that the use of 0.5% of AMP, PVP, Tween-80, and xylene improved the staining ability of protein silver staining, compared with the original method. Furthermore, 0.5% AMP, 0.5% PVP, 0.5% Tween-80 reagents significantly influenced the morphology, size, potential, and dispersion of silver ions. These results suggested a new idea for further improving the detection ability of protein silver staining.
银染是一种检测十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离的蛋白质的极佳技术。与考马斯亮蓝检测法等其他染色技术相比,蛋白质银染技术的灵敏度更高,适用于检测低浓度的蛋白质。本研究旨在提高蛋白质染色方法的检测能力。在此,我们通过添加 AMP、PVP、Tween-80 和二甲苯来改进银染配方,这是一种非常重现性的方法,以增强蛋白质染色的检测能力。此外,使用粒度仪和电位计检测了制备的染色材料中银离子的粒径和电位差,然后使用透射电子显微镜(TEM)研究了不同染色溶液中染色银颗粒的形态、透明度和粒径。结果表明,与原始方法相比,使用 0.5%的 AMP、PVP、Tween-80 和二甲苯可提高蛋白质银染的染色能力。此外,0.5%AMP、0.5%PVP、0.5%Tween-80 试剂显著影响银离子的形态、大小、电位和分散性。这些结果为进一步提高蛋白质银染的检测能力提供了新的思路。