Book contents
- Frontmatter
- Contents
- Preface to the sixth edition
- List of contributors
- List of abbreviations
- 1 Basic principles
- 2 Cell culture techniques
- 3 Centrifugation
- 4 Microscopy
- 5 Molecular biology, bioinformatics and basic techniques
- 6 Recombinant DNA and genetic analysis
- 7 Immunochemical techniques
- 8 Protein structure, purification, characterisation and function analysis
- 9 Mass spectrometric techniques
- 10 Electrophoretic techniques
- 11 Chromatographic techniques
- 12 Spectroscopic techniques: I Atomic and molecular electronic spectroscopy
- 13 Spectroscopic techniques: II Vibrational spectroscopy and electron and nuclear spin orientation in magnetic fields
- 14 Radioisotope techniques
- 15 Enzymes
- 16 Cell membrane receptors
- Index
- Plate sections
10 - Electrophoretic techniques
Published online by Cambridge University Press: 05 June 2012
- Frontmatter
- Contents
- Preface to the sixth edition
- List of contributors
- List of abbreviations
- 1 Basic principles
- 2 Cell culture techniques
- 3 Centrifugation
- 4 Microscopy
- 5 Molecular biology, bioinformatics and basic techniques
- 6 Recombinant DNA and genetic analysis
- 7 Immunochemical techniques
- 8 Protein structure, purification, characterisation and function analysis
- 9 Mass spectrometric techniques
- 10 Electrophoretic techniques
- 11 Chromatographic techniques
- 12 Spectroscopic techniques: I Atomic and molecular electronic spectroscopy
- 13 Spectroscopic techniques: II Vibrational spectroscopy and electron and nuclear spin orientation in magnetic fields
- 14 Radioisotope techniques
- 15 Enzymes
- 16 Cell membrane receptors
- Index
- Plate sections
Summary
GENERAL PRINCIPLES
The term electrophoresis describes the migration of a charged particle under the influence of an electric field. Many important biological molecules, such as amino acids, peptides, proteins, nucleotides and nucleic acids, possess ionisable groups and, therefore, at any given pH, exist in solution as electrically charged species either as cations (+) or anions (-). Under the influence of an electric field these charged particles will migrate either to the cathode or to the anode, depending on the nature of their net charge.
The equipment required for electrophoresis consists basically of two items, a power pack and an electrophoresis unit. Electrophoresis units are available for running either vertical or horizontal gel systems. Vertical slab gel units are commercially available and routinely used to separate proteins in acrylamide gels (Section 10.2). The gel is formed between two glass plates that are clamped together but held apart by plastic spacers. The most commonly used units are the so-called minigel apparatus (Fig. 10.1). Gel dimensions are typically 8.5 cm wide × 5 cm high, with a thickness of 0.5-1 mm. A plastic comb is placed in the gel solution and is removed after polymerisation to provide loading wells for up to 10 samples. When the apparatus is assembled, the lower electrophoresis tank buffer surrounds the gel plates and affords some cooling of the gel plates. A typical horizontal gel system is shown in Fig. 10.2.
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- Information
- Principles and Techniques of Biochemistry and Molecular Biology , pp. 449 - 484Publisher: Cambridge University PressPrint publication year: 2005