The separation of native proteins by electrophoresis using agarose gel as a medium.
The purpose of this experiment was to see which direction certain proteins will migrate according to net charge of protein and pH of buffer used. .
III. Proteins play an important role in the structure and function of all living things. Amino acids are the protein's building blocks. There are 20 amino acids that are found in proteins. What makes them different is the nature of the R-group attached to the alpha carbon. The amino acids are held together by peptide bonds. These bonds help give each protein it's unique structure and function. These three-dimensional shapes are of critical importance to the protein's function. There are three structural levels that define the three-dimensional shape of a protein. The first of these called the secondary structure are the alpha helices and the beta pleated sheets. These structures are held together by hydrogen bonds. The second called the tertiary structure is brought about by the interactions of the alpha helices and the beta pleated sheets as well as any other folds, turns or straight stretches in the molecule. The hydrophobic parts and the hydrophilic parts assume their posit!.
ion so as to either be near the water or away from the water respectively. Lastly is the quaternary structure, which deals with the arrangement of the many polypeptide chains in a variety of noncovalent bonds.
As stated in the beginning, the purpose of this experiment is to use electrophoresis to separate proteins. Electrophoresis is the movement of charged molecules under the influence of an electric field. Since proteins and amino acids are charged molecules, when they are placed into an electric field they will migrate depending on the pH value. The agarose gel has become a regular medium for this type of experiment because it serves as a matrix for the buffer in which the sample molecules will be traveling.