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.
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