SDS-PAGE
SDS-PAGE or Sodium dodecyl sulfate polyacrylamide gel electrophoresis is a technique widely used in biochemistry, forensics, genetics, molecular biology and biotechnology to separate proteins according to their electrophoretic mobility (a function of the length of a polypeptide chain and its charge) and no other physical feature.
Overview[edit | edit source]
SDS-PAGE is a method that enables the separation of molecules by size. The technique is based on the principle that a charged molecule will migrate in an electric field towards an electrode with opposite sign. The rate of migration of a molecule through a gel is inversely proportional to the logarithm of its size. Therefore, smaller molecules will migrate faster and further than larger molecules.
Procedure[edit | edit source]
The procedure of SDS-PAGE can be divided into several steps:
- Sample preparation: The protein sample is mixed with a buffer containing SDS and a reducing agent. The mixture is then heated to denature the proteins.
- Gel preparation: The gel is prepared by polymerizing a mixture of acrylamide and bis-acrylamide.
- Electrophoresis: The samples are loaded into the wells of the gel and an electric current is applied. The proteins migrate towards the positive electrode at a rate proportional to their size.
- Staining: After electrophoresis, the gel is stained to visualize the proteins.
Applications[edit | edit source]
SDS-PAGE is used in various fields for different purposes:
- In biochemistry and molecular biology, it is used to determine the molecular weight of proteins and to analyze their relative abundance in different samples.
- In forensics, it is used to compare samples from crime scenes with samples from suspects.
- In biotechnology, it is used to monitor the purity of protein products.
See also[edit | edit source]
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