2D gel electrophoresis: Difference between revisions
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It is a powerful method of separating [[Protein|protein]] along a tubular gel using [[Isoelectric focusing|iso-electric focusing]] and [[SDS polyacrylamide-gel electrophoresis|sodium dodecyl sulphate polyacrylamide gel]]. The sample undergoes [[Isoelectric focusing|iso-electric focusing]] first; an electrical current is passed through a gel with a pH gradient, and the proteins stop moving when they reach the pH at which they have no net charge. The sample is then placed horizontally on top of [[SDS-PAGE|SDS-PAGE]] and spreads across. The sample then moves vertically down again to yield the second dimension based on the size of the protein, producing a 2D pattern. So the sample has been separated by virtue of [[Isoelectric point|Iso-electric point]] (pI), when they move horizontally and by virtue of their size; when they move down vertically. | It is a powerful method of separating [[Protein|protein]] along a tubular gel using [[Isoelectric focusing|iso-electric focusing]] and [[SDS polyacrylamide-gel electrophoresis|sodium dodecyl sulphate polyacrylamide gel]]. The sample undergoes [[Isoelectric focusing|iso-electric focusing]] first; an electrical current is passed through a gel with a pH gradient, and the proteins stop moving when they reach the pH at which they have no net charge - this is known as the [[Isoelectric_point|Iso-electric point]] (pI). The sample is then placed horizontally on top of [[SDS-PAGE|SDS-PAGE]] and spreads across. The sample then moves vertically down again to yield the second dimension based on the size of the protein, producing a 2D pattern. So the sample has been separated by virtue of [[Isoelectric point|Iso-electric point]] (pI), when they move horizontally and by virtue of their size; when they move down vertically. | ||
Because of the specificity of this method, it is possible to identify over one thousand proteins on the same gel, in one experiment <ref>Stryer, Lubert; Tymoczko, John L.; Berg, Jeremy M., (2012) Biochemistry, 7th Edition, New York: WH Freeman and Co. Page 74</ref>. | |||
=== References === | === References === | ||
<references /> | <references /> | ||
Revision as of 11:53, 29 November 2012
It is a powerful method of separating protein along a tubular gel using iso-electric focusing and sodium dodecyl sulphate polyacrylamide gel. The sample undergoes iso-electric focusing first; an electrical current is passed through a gel with a pH gradient, and the proteins stop moving when they reach the pH at which they have no net charge - this is known as the Iso-electric point (pI). The sample is then placed horizontally on top of SDS-PAGE and spreads across. The sample then moves vertically down again to yield the second dimension based on the size of the protein, producing a 2D pattern. So the sample has been separated by virtue of Iso-electric point (pI), when they move horizontally and by virtue of their size; when they move down vertically.
Because of the specificity of this method, it is possible to identify over one thousand proteins on the same gel, in one experiment [1].
References
- ↑ Stryer, Lubert; Tymoczko, John L.; Berg, Jeremy M., (2012) Biochemistry, 7th Edition, New York: WH Freeman and Co. Page 74