Sds Page (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis)

1326 WordsJun 18, 20126 Pages
Gel electrophoresis is a routine laboratory procedure in biochemical studies that takes advantage of a protein’s amphoteric nature to determine its molecular weight and charge by running the sample through a gel matrix under the influence of an electrically charged field. A popular example of gel electrophoresis is Sodium dodecyl sulfate-polyacrylamide electrophoresis or SDS- PAGE which has been used in this experiment to supposedly determine albumin and casein’s molecular weights respectively. The system used in gel electrophoresis consists of 3 major parts: a stationary phase, a mobile phase) and an electrical power supply. In SDS-PAGE the polyacrylamide gel serves as the stationary phase, the buffer (Tris-HCl in this case) as the…show more content…
On the other hand, the Tris-HCl buffer and deionized water provides the system its shape since the polyacrylamide complex is just a thin film of interlocking polymer fibers without any liquid to trap within. Polyacrylamide gels are usually preferred over other gel types such as agarose in protein electrophoresis because of various benefits such as its ability to be cast in various pore sizes which provides options for the researchers who observe protein samples with various Kda’s depending on how separated they aim the bands to be or how well the sample is sieved. It is also preferred because of its charge neutrality upon setting as it does not interfere with the electrophoresis which is highly reliant on the sample and the surrounding’s charges within the system. Lastly, the gel itself is colorless and protein stains do not adhere to the gel so there will be minimal problems with unnecessary stains that aren’t from the staining solution. APS and TEMED was used to initiate the binding of acrylamide to bisacrylamide via a vinyl addition reaction where TEMED catalyzes the decomposition of APS into free sulfide molecules which then initiates the formation of the polyacrylamide complex. Sodium dodecyl-sulfate or SDS is a protein denaturant that destroys weak interactions within the protein molecule which unfolds it
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