MBSc. Mohammad
04-02-2007, 06:51 AM
Two-dimensional electrophoresis
http://www.moq3.com/pics/up_2007/01_2007/b2f8aab533.jpg
is one of the best experimental tools for the reliable separation of thousands of proteins in a single gel. 2DE consists of a tandem pair of electrophoretic separations
?How does it work
The polyacrylamide gel provides a supporting matrix through which proteins can migrate. This gel has a pH gradient from top to bottom - the top is more acidic than the bottom
A complex protein mixture is loaded in the middle of the left side, where the pH is neutral, and a voltage is applied across the gel. The proteins then migrate through the gel until they reach their isoelectric point . This technique is called isoelectric focussing
The gel is then soaked in a denaturing solution (which causes the proteins to unfold) containing the detergent sodium dodecylsulphate (SDS). This molecule has a very strong negative charge and binds to all proteins, effectively making them all the same charge
A voltage is then applied across the gel from side to side with the anode (positive terminal) at the right. The proteins all move towards the anode, but smaller proteins move faster through the gel than larger ones, thus separating the proteins according to their size
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) is used to separate mixtures of proteins, and is particularly useful for comparing related samples ? such as healthy and diseased tissue. The first step is to load the proteins onto the gel, which has a pH gradient from top to bottom
؟How is it used
2D-PAGE can be used in combination with mass spectrometry to systematically identify all the major proteins present in a sample
However, the technique is particularly powerful when comparing related samples, such as healthy tissue vs disease tissue. For example, proteins that are more abundant in disease tissue may represent novel drug targets or diagnostic disease markers
Comparative 2D-PAGE can also be used to look for proteins whose expression varies similarly under the same set of conditions (these may have related functions) and to identify proteins (produced in response to drug therapy (these may be responsible for drug-related side effects
http://sultan035636.jeeran.com/2d-page.jpg
http://www.moq3.com/pics/up_2007/01_2007/f689847025.jpg
http://sultan035636.jeeran.com/electrophoresis__2d-page_.jpg
read my subject and you know what Two-dimensional electrophoresis :sm198:
http://sultan035636.jeeran.com/electrophoresis_man.jpg
http://www.moq3.com/pics/up_2007/01_2007/b2f8aab533.jpg
is one of the best experimental tools for the reliable separation of thousands of proteins in a single gel. 2DE consists of a tandem pair of electrophoretic separations
?How does it work
The polyacrylamide gel provides a supporting matrix through which proteins can migrate. This gel has a pH gradient from top to bottom - the top is more acidic than the bottom
A complex protein mixture is loaded in the middle of the left side, where the pH is neutral, and a voltage is applied across the gel. The proteins then migrate through the gel until they reach their isoelectric point . This technique is called isoelectric focussing
The gel is then soaked in a denaturing solution (which causes the proteins to unfold) containing the detergent sodium dodecylsulphate (SDS). This molecule has a very strong negative charge and binds to all proteins, effectively making them all the same charge
A voltage is then applied across the gel from side to side with the anode (positive terminal) at the right. The proteins all move towards the anode, but smaller proteins move faster through the gel than larger ones, thus separating the proteins according to their size
Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) is used to separate mixtures of proteins, and is particularly useful for comparing related samples ? such as healthy and diseased tissue. The first step is to load the proteins onto the gel, which has a pH gradient from top to bottom
؟How is it used
2D-PAGE can be used in combination with mass spectrometry to systematically identify all the major proteins present in a sample
However, the technique is particularly powerful when comparing related samples, such as healthy tissue vs disease tissue. For example, proteins that are more abundant in disease tissue may represent novel drug targets or diagnostic disease markers
Comparative 2D-PAGE can also be used to look for proteins whose expression varies similarly under the same set of conditions (these may have related functions) and to identify proteins (produced in response to drug therapy (these may be responsible for drug-related side effects
http://sultan035636.jeeran.com/2d-page.jpg
http://www.moq3.com/pics/up_2007/01_2007/f689847025.jpg
http://sultan035636.jeeran.com/electrophoresis__2d-page_.jpg
read my subject and you know what Two-dimensional electrophoresis :sm198:
http://sultan035636.jeeran.com/electrophoresis_man.jpg