title: A review on preparative and semi-preparative offgel electrophoresis for multidimensional protein/peptide assessment.
abstract: Mass spectrometry (MS) techniques are commonly used for protein identification and further analysis of selected protein spots after high resolution 2-D electrophoresis. Complementary gel-free approaches have been developed during the last few years and have shown to be useful tools in modern proteomics. The development and application of various gel-free electrophoresis devices for performing protein fractionation according to the pI differences is therefore a topic of interest. This review describes the current state of isoelectric focusing (IEF) gel-free electrophoresis based on the Agilent offgel 3100 fractionator. The review includes, therefore, (i) an overview on IEF as well as other previous IEF gel-free electrophoresis developments; (ii) offgel fundamentals and future trends; (iii) advantages and disadvantages of current offgel procedures; (iv) requirements of isolated protein pellets for further offgel fractionation; (v) offgel fraction requirements to perform the second dimensional analysis by advance electrophoresis and chromatographic techniques; and (vi) effect of the offgel operating conditions on the stability of metal-protein complexes.
authors: Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Chemistry, University of Santiago de Compostela, Avenida das Ciencias, s/n. 15782 Santiago de Compostela, Spain. Electronic address: antonio.moreda@usc.es.
authors: Antonio
authors: A
authors: Moreda-Piñeiro
authors: Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Chemistry, University of Santiago de Compostela, Avenida das Ciencias, s/n. 15782 Santiago de Compostela, Spain.
authors: Natalia
authors: N
authors: García-Otero
authors: Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Chemistry, University of Santiago de Compostela, Avenida das Ciencias, s/n. 15782 Santiago de Compostela, Spain.
authors: Pilar
authors: P
authors: Bermejo-Barrera
mesh_terms: D005591:Chemical Fractionation; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods* / Q000639:trends; D007525:Isoelectric Focusing / Q000379:methods; D008670:Metals / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D020033:Protein Isoforms / Q000032:analysis; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000737:chemistry
chemical_list: D008670:Metals; D010455:Peptides; D020033:Protein Isoforms; D011506:Proteins; D014357:Trypsin
keywords: Isoelectric focusing; Offgel electrophoresis; Peptide; Protein