All you need to know about the principle of Two Dimensional (2D) Gel Electrophoresis in Hindi
2D gel electrophoresis (2DE) is a key technique for purifying individual proteins from complex samples based on their isoelectric points and molecular weights.
1.Much like regular SDS-PAGE, those tissue or blood samples for 2DE need to be processed and solubilized before they can be loaded into the IEF gel.
2. Time to load it on an IEF gel where, similar to SDS-PAGE, the proteins will be pushed through the acrylamide gel by an electric field. Where IEF gets more exciting is that the gel incorporates a pH gradient, and each protein moves only until it reaches its isoelectric point (pI). The pI is the pH where a protein has no net charge, meaning the field has no effect and the protein stays.
3..Finally, the second dimension! But first, proteins need to be solubilized again in SDS before they can be separated by their molecular weights on an second axis. Other than this equilibration and some additional care with timing and voltages, this step uses molecular weights to separate proteins through the tried-and-true SDS-PAGE method. At the end, the gel will have the proteins aligned along two axes: isoelectric point vs. molecular weight.
Please watch this video to have a clear idea about 2D gel electrophoresis.
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SDS PAGE | Part 1 - Gel Preparation | Hindi : https://youtu.be/vepxeDep6fk
SDS PAGE | Part 2 - Principle | Hindi : https://youtu.be/Tw5rGCNBlR0
Gel Electrophoresis | Agarose Gel Electrophoresis Lab Procedure : https://youtu.be/citMv0hvfyc
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