College of Chemistry, Beijing Normal University, Beijing 100875, P.R. China.
Chemistry. 2012 Jan 27;18(5):1432-7. doi: 10.1002/chem.201101310. Epub 2012 Jan 16.
We have developed a novel application for DNA oligonucleotide-stabilized Ag nanoclusters in fluorescent imaging of human serum proteins after native polyacrylamide gel electrophoresis (PAGE). Oligonucleotide-stabilized Ag nanoclusters were used as fluorescent probes for direct detection of proteins after native PAGE. Some relatively low-abundance proteins, such as α-1-antichymotrypsin (ACT) and α-2-glycoprotein 1, zinc (ZAG) were easily detected by oligonucleotide-stabilized Ag nanocluster-based fluorescent imaging and identified by MS and MS/MS techniques, without the need of expensive antibodies or tedious immunoassay procedures. The pH condition for the oligonucleotide-stabilized Ag nanocluster solution was optimized and the possible mechanism of interaction between proteins and DNA oligonucleotide-stabilized Ag nanoclusters was analyzed. As a novel fluorescent detection method it is simple, fast, nontoxic and sensitive, and it shows great analytical potential in proteome research and in biochemistry.
我们开发了一种新的应用,将 DNA 寡核苷酸稳定的银纳米簇应用于未经处理的聚丙烯酰胺凝胶电泳(PAGE)后人血清蛋白质的荧光成像。寡核苷酸稳定的银纳米簇被用作荧光探针,用于直接检测未经处理的 PAGE 后的蛋白质。一些相对低丰度的蛋白质,如α-1-抗胰蛋白酶(ACT)和α-2-糖蛋白 1,锌(ZAG),可以通过基于寡核苷酸稳定的银纳米簇的荧光成像轻松检测,并通过 MS 和 MS/MS 技术进行鉴定,而无需昂贵的抗体或繁琐的免疫测定程序。优化了寡核苷酸稳定的银纳米簇溶液的 pH 值条件,并分析了蛋白质与 DNA 寡核苷酸稳定的银纳米簇之间相互作用的可能机制。作为一种新型的荧光检测方法,它简单、快速、无毒且灵敏,在蛋白质组学研究和生物化学领域具有巨大的分析潜力。