Cryogenic Postseparation Treatment of Acrylamide Gels for Metaloproteomic Analyses by Combination of Gel Electrophoresis, Laser Ablation and Inductively Coupled Plasma Mass Spectrometry

Authors

  • Hana Ovadová-Nejezchlebová Masaryk University, Faculty of Science, Department of Experimental Biologiy, Division of Functional Genomics and Proteomics, Central Laboratory, Brno, Czech Republic
  • Tomáš Vaculovič Masaryk University, Central-European Institute of Technology, Centre for Nanotechnology and Microtechnology, Laboratory for Synthesis and Analysis of Nanostructures, Brno, Czech Republic
  • Paul Ek Abo Akademi University, Laboratory of Analytical Chemistry, Abo (Turku), Finland
  • Jan Havliš Masaryk University, Central-European Institute of Technology, Mendel Centre for Plant Genomics and Proteomics, Proteomics CF, Brno, Czech Republic | Masaryk University, Faculty of Science, Department of Experimental Biologiy, Division of Functional Genomics and Proteomics, Central Laboratory, Brno, Czech Republic
  • Viktor Kanický Masaryk University, Faculty of Science, Department of Chemistry, Divisionof Analytical Chemistry, Laboratory of Atomic Spectrochemistry, Brno, Czech Republic | Masaryk University, Central-European Institute of Technology, Centre for Nanotechnology and Microtechnology, Laboratory for Synthesis and Analysis of Nanostructures, Brno, Czech Republic

Keywords:

gel electrophoresis, laser ablation, inductively coupled plasma, mass spectrometry, metalloprotein, metal-binding protein, cryogenic ablation cell, cellophane film, gel drying

Abstract

Analysis of metal species in cryogenic frozen gels originated in polyacrylamide gel electrophoresis (PAGE) by laser ablation (LA) connected to inductively coupled plasma mass spectrometry (ICP-MS) – GLIM in abbreviation – has been investigated to detect/quantitate them in separated metalloproteins. It was compared to traditional ways of gel drying procedures prior to ablation (plain drying, drying gel sandwiched between two cellophane foils), which are necessary operations before laser ablation to prevent undesired interferences related to gas production during laser ablation because of liquid content. Laser ablation of the gels containing BSA:Co(II) 1:1 and ADH with Zn(II) in cryogenic ablation cell (at –80 °C for 25 min) is a relevant alternative to the traditional procedures because of significant time spared during the procedure (down to tens of hours instead of days), it yields comparable outcome of metal species analysis by means of their high signal intensities and low RSD in ICP-MS and because of reusability of gels after cryogenic ablation, e.g. for mass spectrometry driven identification of protein moiety of metalloprotein.

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Published

2011-11-15

How to Cite

Ovadová-Nejezchlebová, H., Vaculovič, T., Ek, P., Havliš, J., & Kanický, V. (2011). Cryogenic Postseparation Treatment of Acrylamide Gels for Metaloproteomic Analyses by Combination of Gel Electrophoresis, Laser Ablation and Inductively Coupled Plasma Mass Spectrometry. Chemické Listy, 105(11), 896–902. Retrieved from http://blog.chemicke-listy.cz/ojs3/index.php/chemicke-listy/article/view/1036

Issue

Vol. 105 No. 11 (2011)

Section

Articles

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