changes in protein profiles in bortezomib applied multiple myeloma ...
changes in protein profiles in bortezomib applied multiple myeloma ...
changes in protein profiles in bortezomib applied multiple myeloma ...
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2.1.4.1. Gel Electrophoresis<br />
Electrophoresis is a bio-analytical method utilized for the isolation of<br />
biomolecules depend<strong>in</strong>g on the mobility of charged molecules under the <strong>in</strong>fluence of an<br />
electric field. When an electric field is <strong>applied</strong>, any charged molecule <strong>in</strong> solution will<br />
migrate. (Twyman, 2004). Mobility of a molecule is based upon the magnitude of its<br />
charge, molecular weight, and structure. Many biopolymers, such as prote<strong>in</strong>s are<br />
charged either by coat<strong>in</strong>g with the anionic detergent sodium dodecylsulfate (SDS) or by<br />
acid-base association-dissociation reactions of am<strong>in</strong>e and carboxylic acid parts of them,<br />
so they can also be separated by electrophoretic methods (Mikkelsen and Corton 2004).<br />
Electrophoresis, which is the central component of the proteomic research, is<br />
commonly carried out <strong>in</strong> a gel that is formed by the polymerization of acrylamide, so it<br />
is called polyacrylamide gel electrophoresis. The reaction mechanism that results <strong>in</strong> a<br />
gel with a characteristic porosity which depends on the polymerization conditions and<br />
monomer concentrations can be seen <strong>in</strong> Figure 2.3 (K<strong>in</strong>ter and Sherman 2000).<br />
Figure 2.3. Reaction of polyacrylamide gel formation<br />
(Source: Gallagher, 2008)<br />
In the presence of an <strong>in</strong>itiator generally ammonium persulfate and a catalyst<br />
commonly tetramethylethylenediam<strong>in</strong>e [TEMED, (CH3)2N(CH2)2N(CH3)2],<br />
polyacrylamide gels are prepared by the copolymerization of acrylamide monomer with<br />
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