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SDS-PAGE as a Tool for Hydrodynamic Diameter-Dependent Separation of Quantum Dots

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Abstract

The purpose of this study was to test quantum dots (QDs) separation by native and Tris–Glycine SDS-PAGE according to the protocols commonly used for protein analyses. To study the electrophoretic behaviour of quantum dots, ten samples of previously synthesized CdTe QDs stabilized with mercaptosuccinic acid (MSA) were used. Prior to electrophoresis the hydrodynamic diameters of QDs and zeta potentials were determined, as well as the fluorescence properties and stability of QDs in the running buffers. After verification of QDs stability and separation in native polyacrylamide gel, SDS-PAGE in gradient 4–20 % polyacrylamide gel was performed. Under UV irradiation a colour-dependent separation of QDs was observed, which was consistent with their hydrodynamic diameter distribution. The electrophoretic conditions were further optimized with respect to achieving the optimal colour separation, fluorescence stability and to minimize the time of analysis. Based on the results obtained, for further work 15 % polyacrylamide gels with SDS were used and the times (30–60 min) and voltage (100–150 V) used for separation were optimized. Under the optimal separation conditions (30 min, 100 V) the addition of MSA in the concentration range 0–4 mM was used to improve visualization of QDs with diameters in the range from 7 ± 2 to 4 ± 2 nm.

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Abbreviations

QDs:

Quantum dots

DLS:

Dynamic light scattering

AGE:

Agarose gel electrophoresis

TAE:

Tris-acetate-EDTA

TBE:

Tris-borate-EDTA

PAGE:

Polyacrylamide gel electrophoresis

DNA:

Deoxyribonucleic acid

SDS:

Sodium-dodecyl sulphate

SDS-PAGE:

Polyacrylamide gel electrophoresis in presence of sodium-dodecyl sulphate

MPA:

Mercaptopropionic acid

MSA:

Mercaptosuccinic acid

T-G:

Tris-glycine

EDTA:

Ethylenediaminetetraacetic acid

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Acknowledgments

Financial support from NanoBioTECell GA CR P102/11/1068 is greatly acknowledged. Authors would like to thank to Ms. Dagmar Uhlirova for technical assistance.

Conflict of interest

The authors have declared no conflict of interest.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, 613 00, Brno, Czech Republic

    Sona Krizkova, Simona Dostalova, Petr Michalek, Lukas Nejdl, Marketa Kominkova, Vedran Milosavljevic, Amitava Moulick, Marketa Vaculovicova, Pavel Kopel, Vojtech Adam & Rene Kizek

  2. Central European Institute of Technology, Brno University of Technology, Technicka 3058/10, 616 00, Brno, Czech Republic

    Sona Krizkova, Simona Dostalova, Petr Michalek, Lukas Nejdl, Marketa Kominkova, Vedran Milosavljevic, Amitava Moulick, Marketa Vaculovicova, Pavel Kopel, Vojtech Adam & Rene Kizek

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  1. Sona Krizkova
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  2. Simona Dostalova
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Correspondence to Rene Kizek.

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Krizkova, S., Dostalova, S., Michalek, P. et al. SDS-PAGE as a Tool for Hydrodynamic Diameter-Dependent Separation of Quantum Dots. Chromatographia 78, 785–793 (2015). https://doi.org/10.1007/s10337-015-2893-z

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  • DOI: https://doi.org/10.1007/s10337-015-2893-z

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