Journal
ACS NANO
Volume 3, Issue 6, Pages 1318-1328Publisher
AMER CHEMICAL SOC
DOI: 10.1021/nn9000353
Keywords
Western blot; quantum dot; electrophoresis; single molecule; proteomics; immunoblot
Categories
Funding
- DOD [W81 XWH-07-2-0107]
- Oregon ETIC funds
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Substantially improved detection methods are needed to detect fractionated protein samples present at trace concentrations in complex, heterogeneous tissue and biofluid samples. Here we describe a modification of traditional Western immunoblotting using a technique to count quantum-dot-tagged proteins on optically transparent PVDF membranes. Counts of quantum-dot-tagged proteins on immunoblots achieved optimal detection sensitivity of 0.2 pg and a sample size of 100 cells. This translates to a 10(3)-fold improvement in detection sensitivity and a 10(2)-fold reduction in required cell sample, compared to traditional Westerns processed using the same membrane immunoblots. Quantum dot fluorescent blinking analysis showed that detection of single QD-tagged proteins is possible and that detected points of fluorescence consist of one or a few (< 9) QDs. The application of single nanoparticle detection capabilities to Western blotting technologies may provide a new solution to a broad range of applications currently limited by insufficient detection sensitivity and/or sample availability.
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