TITLE

A simple, sensitive and selective quantum-dot-based western blot method for the simultaneous detection of multiple targets from cell lysates

AUTHOR(S)
Gilroy, Kathryn L.; Cumming, Sarah A.; Pitt, Andrew R.
PUB. DATE
September 2010
SOURCE
Analytical & Bioanalytical Chemistry;Sep2010, Vol. 398 Issue 1, p547
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Quantum dots (Qdots) are fluorescent nanoparticles that have great potential as detection agents in biological applications. Their optical properties, including photostability and narrow, symmetrical emission bands with large Stokes shifts, and the potential for multiplexing of many different colours, give them significant advantages over traditionally used fluorescent dyes. Here, we report the straightforward generation of stable, covalent quantum dot–protein A/G bioconjugates that will be able to bind to almost any IgG antibody, and therefore can be used in many applications. An additional advantage is that the requirement for a secondary antibody is removed, simplifying experimental design. To demonstrate their use, we show their application in multiplexed western blotting. The sensitivity of Qdot conjugates is found to be superior to fluorescent dyes, and comparable to, or potentially better than, enhanced chemiluminescence. We show a true biological validation using a four-colour multiplexed western blot against a complex cell lysate background, and have significantly improved previously reported non-specific binding of the Qdots to cellular proteins. [Figure not available: see fulltext.]
ACCESSION #
52815217

 

Related Articles

  • New opportunities in multiplexed optical bioanalyses using quantum dots and donor-acceptor interactions. Algar, W. Russ; Krull, Ulrich J. // Analytical & Bioanalytical Chemistry;Nov2010, Vol. 398 Issue 6, p2439 

    This review highlights recent trends in the development of multiplexed bioanalyses using quantum dot bioconjugates and donor-acceptor interactions. In these methods, multiple optical signals are generated in response to biorecognition through modulation of the photoluminescence of populations of...

  • Imaging: In vivo tracking. Pile, David // Nature Photonics;Apr2013, Vol. 7 Issue 4, p262 

    The article focuses on the study of the role of quantum dots or fluorosphores in tracking protein locations and other species in a biological cells is presented.

  • Fluorescence-Based Multiplex Protein Detection Using Optically Encoded Microbeads. Bong-Hyun Jun; Homan Kang; Yoon-Sik Lee; Dae Hong Jeong // Molecules;Mar2012, Vol. 17 Issue 3, p2474 

    Potential utilization of proteins for early detection and diagnosis of various diseases has drawn considerable interest in the development of protein-based multiplex detection techniques. Among the various techniques for high-throughput protein screening, optically-encoded beads combined with...

  • Host-Guest Carbon Dots for Enhanced Optical Properties and Beyond. Sun, Ya-Ping; Wang, Ping; Lu, Zhuomin; Yang, Fan; Meziani, Mohammed J.; LeCroy, Gregory E.; Liu, Yun; Qian, Haijun // Scientific Reports;7/24/2015, p12354 

    Carbon dots, generally small carbon nanoparticles with various forms of surface passivation, have achieved the performance level of semiconductor quantum dots in the green spectral region, but their absorption and fluorescence in red/near-IR are relatively weaker. Conceptually similar to...

  • Semiconductor Nanomaterials-Based Fluorescence Spectroscopic and Matrix-Assisted Laser Desorption/Ionization (MALDI) Mass Spectrometric Approaches to Proteome Analysis. Kailasa, Suresh Kumar; Kuang-Hung Cheng; Hui-Fen Wu // Materials (1996-1944);Dec2013, Vol. 6 Issue 12, p5763 

    Semiconductor quantum dots (QDs) or nanoparticles (NPs) exhibit very unusual physico-chemcial and optical properties. This review article introduces the applications of semiconductor nanomaterials (NMs) in fluorescence spectroscopy and matrix-assisted laser desorption/ionization mass...

  • Multiplexed measurements by time resolved spectroscopy using colloidal CdSe/ZnS quantum dots. Kaiser, U.; Jimenez de Aberasturi, D.; Malinowski, R.; Amin, F.; Parak, W. J.; Heimbrodt, W. // Applied Physics Letters;1/27/2014, Vol. 104 Issue 4, p041901-1 

    Multiplexed measurements of analytes in parallel is a topical demand in bioanalysis and bioimaging. An interesting alternative to commonly performed spectral multiplexing is lifetime multiplexing. In this Letter, we present a proof of principle of single-color lifetime multiplexing by coupling...

  • Optical properties of lnAs quantum dots in a Si matrix. Heitz, R.; Ledenstov, N.N. // Applied Physics Letters;3/22/1999, Vol. 74 Issue 12, p1701 

    Investigates the optical properties of quantum dots in a silicon matrix. Growth temperature; Electronic structure of quantum dots; Excitation density dependence of the integrated quantum dot.

  • Can nanoparticles be useful for antiviral therapy? Lozovski, V.; Lysenko, V.; Pyatnitsia, V.; Spivak, M. // Semiconductor Physics, Quantum Electronics & Optoelectronics;2011, Vol. 14 Issue 4, p489 

    In this paper, optical properties of the system consisting of mesoparticles (small dielectric particles) and nanoparticles (quantum dots) of various shapes have been considered. This system can be characterized by resonant absorption of electromagnetic waves and used for developing the new...

  • Photoluminescence decay rate engineering of CdSe quantum dots in ensemble arrays embedded with gold nano-antennae. Haridas, M.; Basu, J. K.; Tiwari, A. K.; Venkatapathi, M. // Journal of Applied Physics;Aug2013, Vol. 114 Issue 6, p064305 

    We discuss experimental results on the ability to significantly tune the photoluminescence decay rates of CdSe quantum dots embedded in an ordered template, using lightly doped small gold nanoparticles (nano-antennae), of relatively low optical efficiency. We observe both enhancement and...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics