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Modern Bioanalysis of Proteins by Electrophoretic Techniques

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Protein Downstream Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1129))

Abstract

In 1957, protein rich in cysteine able to bind cadmium was isolated from horse kidney and named as metallothionein according to its structural properties. Further, this protein and metallothionein-like proteins have been found in tissues of other animal species, yeasts, fungi and plants. MT is as a potential cancer marker in the focus of interest, and its properties, functions, and behavior under various conditions are intensively studied. Our protocol describes separation of two major mammalian isoforms of MT (MT-1 and MT-2) using capillary electrophoresis (CE) coupled with UV detector. This protocol enables separation of MT isoforms and studying of their basic behavior as well as their quantification with detection limit in units of ng per μL. Sodium borate buffer (20 mM, pH 9.5) was optimized as a background electrolyte, and the separation was carried out in fused silica capillary with internal diameter of 75 μm and electric field intensity of 350 V/cm. Optimal detection wavelength was 254 nm.

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Acknowledgements

Financial support from CEITEC CZ.1.05/1.1.00/02.0068 is highly acknowledged.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University, Brno, Czech Republic

    Sona Krizkova, Marketa Ryvolova, Michal Masarik, Ondrej Zitka, Vojtech Adam & Jaromir Hubalek

  2. Central European Institute of Technology, Brno University of Technology, Brno, Czech Republic

    Sona Krizkova, Marketa Ryvolova, Michal Masarik, Ondrej Zitka, Vojtech Adam & Jaromir Hubalek

  3. Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic

    Michal Masarik

  4. Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine, Charles University, Prague, Czech Republic

    Tomas Eckschlager

  5. University Hospital Motol, Prague, Czech Republic

    Tomas Eckschlager

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

    Rene Kizek

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  1. Sona Krizkova
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  2. Marketa Ryvolova
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Correspondence to Rene Kizek .

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Editors and Affiliations

  1. Laboratory of Enzyme Technology, Agricultural University of Athens Department of Agr. Biotechnology, Athens, Greece

    Nikolaos E. Labrou

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Krizkova, S. et al. (2014). Modern Bioanalysis of Proteins by Electrophoretic Techniques. In: Labrou, N. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 1129. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-977-2_28

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  • DOI: https://doi.org/10.1007/978-1-62703-977-2_28

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