Developing a sensitive and quantitative western blot kit

摘要 蛋白质印迹技术是目前应用最广泛的检测目标蛋白质的生物分析技术之一。 在未知的混合物中，如细胞/组织匀浆或提取物。这项技术是在晚期发明的。 20世纪70年代，在过去的40年里，他在生物学和医学方面取得了许多发现。尽管它不是 在聚光灯下，Western blotting的使用频率至少是其他竞争对手的两倍 或补充技术，如质谱仪，酶联免疫吸附试验和IHC在过去10年。尽管如此 令人惊讶的是，科学界广泛接受了西方印迹法，总的来说，它不是一种非常敏感的方法。 技术，只是半定量的。蛋白质印迹中缺乏定量已经引起了人们对 科学界对数据的可重复性进行了研究。在我们近年来取得的两项突破的基础上， 单价量子点探针和光学透明的转移膜，我们建议开发下一代 Western印迹试剂盒具有更高的灵敏度、量化能力、多路传输能力和读出灵活性 (标准凝胶UV成像仪或基于显微镜的颗粒计数)。如果成功，我们预计这项技术将 迅速成为一种商业产品，为下一个40年的生物学研究提供动力。 。

ABSTRACT Western blotting is one of the most widely used bioanalytical techniques for the detection of proteins of interest in an unknown mixture such as cell/tissue homogenate or extract. This technology was invented in the late 1970s and has enabled numerous discoveries in biology and medicine in the past 40 years. Although it is not in the spotlight anymore, western blotting has been applied at least two times more often than other competing or complementing techniques, such as mass spec, ELISA, and IHC within the last 10 years. Despite this amazingly wide acceptance by the scientific community, western blotting, in general, is not a very sensitive technique and is only semi-quantitative. The lack of quantitation in Western blot has raised major concerns in the scientific community on data reproducibility. Building on two breakthroughs we made in recent years, monovalent Qdot probes and optically clear transfer membranes, we propose to develop the next generation Western blot kits with improved sensitivity, quantification ability, multiplexing capability and flexibility in readout (standard gel UV imager or microscopy-based particle counting). If successful, we expect this technology to quickly become a commercial product to power biology research for another 40 years. .