2-DE

1. TWO –DIMENSIONAL
PROTEIN ELECTROPHORESIS
By,
RAHUL GHALME
B.Tech. (Chemical Engineering)

2. Introduction
 Introduction by O'Farrell and Klose in
1975
 Also known as
◦ 2-DE
◦ 2-D electrophoresis
◦ Two dimensional gel electrophoresis.
 Excellent resolution
 Form of gel electrophoresis
 Separation of mixed proteins

3. Basis for Separation
 Series combination of two electrophoretic
separations
 Based on
1. Charge, using isoelectric focusing
2. Size

4. Procedure
 Place the sample in gel with pH gradient
 Apply potential difference across it

5. Detection Of Proteins
 Using chemical stains like
◦ Silver
◦ Coomassie Brilliant Blue
 2D gel analysis software
◦ BioNumerics 2D, Delta2D,
◦ ImageMaster
◦ PDQuest, Progenesis,
◦ Melanie

6. Uses
 Analyze proteins
 Separation of proteins
 Excellent resolution
 To determine amino acid sequence
 Separation of DNA

7. Thank You