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Polyacrylamide Gel Electrophoresis

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Sodium dodecyl sulfate polyacrylamide gel electrophoresis also known as SDS-PAGE is one of the methods for determining the molecular weight of unknown proteins. SDS is an anionic molecule which denaturizes proteins and brings it back to its’ primary structure and it also provides a negative charge to the uncharged molecule. The SDS-PAGE enables the separation of proteins based on their sizes. The larger the size of the protein, the harder it is to travel through the gel thus heavier proteins stay near the cathode side of the gel. For this experiment, a software named Gel Analyzer was used in order to obtain the molecular weight of the unknown proteins with the help of a protein ladder with known molecular weight and protein concentration. The software also generated a curve with a coefficient determination (r2) of the fit which is equal to 0.998. An equation was also generated from the software to be able to determine the molecular weight of the unknown proteins. Results obtained show that the proteins of the cellular fractions of the chicken liver …show more content…

The gel was submerged into Coomasie blue in order for the stain to bind with the proteins and was again destained afterwards to remove excess proteins. Figure 1.0 shows the results obtained from the SDS PAGE of the cellular fraction of a chicken liver after destaining. The software Gel Analyzer was used in order to determine the lanes, the bands and the molecular weight of each protein. The lanes found in the gel determine the different cellular fractions. The 1st lane found in the left most side is the protein ladder, which has known molecular weight and functions as the ‘ruler’ for the molecular weight of the unknowns. The 2nd band represents the microsome, followed by the crude then another microsome. The 5th band is the nuclei and the 6th and 7th bands are the replicates of the soluble and the nuclei

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