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; | 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 <ref>Berg, J. Stryer, L. Tymoczko, J. (2011) Biochemistry, 7th Edition, New York: W.H Freeman and Company. Chapter 3, Page 76.</ref>. | ||
=== References === | === References === | ||
<references /> | <references /> | ||
Revision as of 15:51, 16 October 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. 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 [1].
References
- ↑ Berg, J. Stryer, L. Tymoczko, J. (2011) Biochemistry, 7th Edition, New York: W.H Freeman and Company. Chapter 3, Page 76.