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2D-SDS Gel Electrophoresis

2D-SDS Gel Electrophoresis

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2D-SDS Gel Electrophoresis

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  1. 2D-SDS Gel Electrophoresis • July 1, 2003 • Theoretical basis for 2D-SDS Gel Elect • The SWISS-2DPAGE Database • Workshop

  2. 2D-SDS Gel ElectrophoresisWhat is it? • This is a method for the separation and identification of proteins in a sample by displacement in 2 dimensions oriented at right angles to one another. • This allows the sample to separate over a larger area, increasing the resolution of each component.

  3. First dimension • Separate by isoelectric point • Isoelectric point-pH at which the net charge on molecule is zero

  4. Visuals http://www.aber.ac.uk/parasitology/Proteome /Tut_2D.html#Section%203

  5. Figure 4-44. Alberts et al. Molecular Biology of the Cell, 3rd edn.

  6. 2nd Dimension-to separate by size • Five steps: • 1. Isolate proteins • 2. Denature proteins by adding sodium dodecyl sulfate • 3. Load onto gel • 4. Apply voltage gradient • 5. Stain or blot

  7. http://www.davidson.edu/academic/biology /courses/Molbio/SDSPAGE/SDSPAGE.html

  8. http://www.davidson.edu/academic/biology /courses/Molbio/SDSPAGE/SDSPAGE.html

  9. http://www.davidson.edu/academic/biology /courses/Molbio/SDSPAGE/SDSPAGE.html

  10. Figure 4-44. Alberts et al. Molecular Biology of the Cell, 3rd edn.

  11. Figure 3-42. Lodish et al. Molecular Cell Biology.

  12. http://www.weihenstephan.de/blm/deg/ manual/manualwork2html02testp9.htm

  13. Databases of 2D-Gels http://www-lmmb.ncifcrf.gov/EP/table2Ddatabases.html http://www.umh.ac.be/~biochim/balhf.htm