2D-SDS Gel Electrophoresis

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2D-SDS Gel Electrophoresis

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

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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.

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First dimension

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

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Visuals
http//www.aber.ac.uk/parasitology/Proteome /Tut_2
D.htmlSection203
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Figure 4-44. Alberts et al. Molecular Biology of
the Cell, 3rd edn.
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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

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http//www.davidson.edu/academic/biology /courses/
Molbio/SDSPAGE/SDSPAGE.html
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http//www.davidson.edu/academic/biology /courses/
Molbio/SDSPAGE/SDSPAGE.html
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http//www.davidson.edu/academic/biology /courses/
Molbio/SDSPAGE/SDSPAGE.html
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Figure 4-44. Alberts et al. Molecular Biology of
the Cell, 3rd edn.
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Figure 3-42. Lodish et al. Molecular Cell
Biology.
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http//www.weihenstephan.de/blm/deg/ manual/manual
work2html02testp9.htm
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Databases of 2D-Gels
http//www-lmmb.ncifcrf.gov/EP/table2Ddatabases.ht
ml
http//www.umh.ac.be/biochim/balhf.htm
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