The overall goal of cellular fractionation is to isolate different cellular components from each other, while still preserving each components function. This experiment consisted of three parts, cellular fractionation (2A), protein quantification of cellular fractions (2B), and enzyme markers to establish fractional purity (2C). The purpose of experiment 2A was to isolate bulk quantities of organelles and other cellular components such as the nuclei, cytosol, and mitochondria. Once these are isolated, the functioning of cellular constituents can more easily be studied. In order to do this, the technique of cellular fractionation is applied. Fractionation begins with homogenization, which involves the break down of cells and the disruption of the cell membranes by shearing forces with liquids, ultra sound, and osmotic shock. If you continuously centrifuge your samples and steadily higher speeds, you will be able to fractionate different parts of a cell into their individual …show more content…
Quantifying this protein content can help scientist later research biochemical activity of the cells. In this method the Coomassie Blue binds to proteins to form a blue complex whose absorbance is directly proportional to protein concentration. The protein content was measured in micrograms per milliliter. Using the protein content of each cellular fraction a standard curve was constructed to show the differences in absorbance. Making serial dilutions of Bovine Serum Albumen (BSA) allowed for the construction of the standard curve. BSA will dissolve in water to form a colorless solution. The dissolved BSA will react with Coomassie Blue to turn solutions into various shades of blue depending on the concentration of protein present in each sample. The higher the protein concentration, the deeper blue the solution will