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Capillary Electrophoretic Enzyme Assays

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Book cover Capillary Electrophoresis of Biomolecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 984))

Abstract

In the past years, capillary electrophoresis has become a frequently used technique for enzyme assays due to the high separation efficiency and versatility as well as small sample size and low consumption of chemicals. The capillary electrophoresis assays can be divided into two general categories: pre-capillary (or offline) assays and in-capillary (or online) assays. In pre-capillary assays, the incubation is performed offline and substrate(s) and product(s) are subsequently analyzed by capillary electrophoresis. In in-capillary assays enzyme reaction and separation of the analytes are performed inside the same capillary. In such assays the enzyme is either immobilized or in solution. The latter techniques is also referred to as electrophoretically mediated microanalysis (EMMA) indicating that the individual steps of the incubation as well as analysis are performed via electrophoretic phenomena. This chapter describes both techniques using the deacetylation of acetyl-lysine residues in model peptides by sirtuin enzymes as well as the hydrolysis of acetylthiocholine by acetylcholinesterase as examples.

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Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, School of Pharmacy, Friedrich Schiller University, Jena, Germany

    Gerhard K. E. Scriba & Hans Abromeit

  2. Department of Pharmaceutical Chemistry, School of Pharmacy, University of Jena, Jena, Germany

    Martina Hense

  3. Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, USA

    Yi Fan

Authors
  1. Gerhard K. E. Scriba
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  2. Hans Abromeit
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  3. Martina Hense
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  4. Yi Fan
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Corresponding author

Correspondence to Gerhard K. E. Scriba .

Editor information

Editors and Affiliations

  1. Department of Life Sciences, University of Modena and Reggio Emilia, Via Campi 213/D, Modena, 41125, Italy

    Nicola Volpi

  2. Department of Life Sciences, University of Modena and Reggio Emilia, Via Campi 213/D, Modena, 41125, Italy

    Francesca Maccari

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Scriba, G.K.E., Abromeit, H., Hense, M., Fan, Y. (2013). Capillary Electrophoretic Enzyme Assays. In: Volpi, N., Maccari, F. (eds) Capillary Electrophoresis of Biomolecules. Methods in Molecular Biology, vol 984. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-296-4_21

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  • DOI: https://doi.org/10.1007/978-1-62703-296-4_21

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-295-7

  • Online ISBN: 978-1-62703-296-4

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