Two Dimensional Polyacrylamide gel Electrophoresis (2D-PAGE) is a technique used to separate and identify proteins in a biological sample. It involves two sequential steps - isoelectric focusing and SDS-PAGE - to separate proteins based on their isoelectric point and molecular weight. This allows visualization of up to 1,000s of individual protein spots on the gel. The protein spots can then be analyzed through techniques like mass spectrometry to identify specific proteins. While 2D-PAGE provides high resolution of complex protein mixtures, it has limitations such as a narrow dynamic range and difficulties separating some classes of proteins.

1. Two Dimensional
Polyacrylamide gel
Electrophoresis (2D-PAGE)
Supervised by-
Dr. Suhas Sutar Bharat
Presented by- IPSITA SAHOO
Reg. no- 200705180160
DOMAIN-
Genomics And Genetic
Engineering

2. CONTENTS
Introduction; 2D PAGE
Workflow of 2D PAGE
1D; Isoelectric focusing
2D; SDS-PAGE
Visualization of results; staining
Advantages vs. Disadvantages
Reference

3.  2D-PAGE stands for Two Dimensional Polyacrylamide gel Electrophoresis
 2D-PAGE is a form of gel electrophoresis in which separation and identification of
proteins in a sample are done by displacement in 2 dimensions oriented at right angles
to one another(orthogonal).
 2D-PAGE is a widely used method for the analysis of complex protein mixture
extracted from cells, tissues, or other biological samples.
 This techniques was first developed by O’Farrell and Klose in 1975.
 2D-PAGE is used in 3 sequential steps-

4. Work flow of 2D

7. 1.
2.

8. Using 2D-PAGE,100 –1000’s of
polypeptides can be analyzed in
a single run.
The proteins can be separated in
pure form from the spots. The
spots can be quantified and also
analyzed by MS.
In this method polypeptides can
be probed with antibodies and
also they can be tested for post-
translational modifications.
2D-PAGE has limited dynamic
range.
This technique include a large
amount of sample handling, less
reproducibility.
It is also not automated for high
throughput analysis.
Difficulty to separate low
abundance proteins, acidic and
basic proteins, very large and
very small proteins and
hydrophobic proteins.
Advantages and Disadvantages

9. References
Beranova-Giorgianni, S. (2003) Proteome analysis by two-dimensional gel
electrophoresis and mass spectrometry: strengths and limitations. TrAC Trends in
Analytical Chemistry 22(5), 273-281.
Stephen J Fey* and Peter Mose Larsen(2001)2D or not 2D. Current Opinion in
Chemical Biology 2001, 5:26–33.
Wayne F Patton*, Birte Schulenberg and Thomas H Steinberg(2002) Two-dimensional
gel electrophoresis; better than a poke in the ICAT?. Current Opinion in Biotechnology,
13:321–328.
David E. Garfin(2003)Two-dimensional gel electrophoresis: an overview.Trends in
Analytical Chemistry, Vol. 22, No. 5
http://www.weihenstephan.de/blm/deg/manual/manualwork2html02test.htm
http://www.aber.ac.uk/parasitology/Proteome/Tut_2D.html