Developing a sensitive and quantitative western blot kit

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. .NARRATIVE Invented 40 years ago, western blotting is one of the most frequently used analytical techniques in biology but suffers from low sensitivity and quantification capability. These limitations are well-recognized by the scientific community but without much effort to address them, because the technology development phase for western blotting has long-passed. Although not in the spot light anymore, the need for a sensitive and quantitative western blot remains strong in the era of quantitative biology. Here, we propose to develop such a kit that can quickly become a commercial product by combining an optically clear transfer membrane and a bright monovalent imaging probe.