Blue Dextran Lab Report

Table 1 mentions the Column volumes of Gel Filtration experiment: Void Volume (Vo), Elution Volume (Ve), Included Volume (Vi), Entire Volume (Vo + Vi), and Inert Volume (Vg). Void Volume (Vo) came out to be 1.25 ml. It was calculated by adding all the fractions that contained blue dextran. Blue Dextran was the biggest substance in size which made it impossible to fit in the available pore sizes of the beads. Void Volume represents the space between the beads in column. In addition, Elution Volume (Ve) came out to be 5.00 ml. It was calculated by adding all of the fractions from beginning to the last fraction that contained yellow food dye. Yellow food dye was the smallest substance in size which made it easy for it to fit in all the pore sizes

In addition, substances collected in fractions were fairly separated because different colors were noticed as it is seen in Table 2: Gel Filtration Fractions. Fraction 4 contained light blue color which suggested the presence of Blue Dextran but in results it was concluded that Fraction 4 did not contain protein. Thus Fraction 5 which contained Dark Blue color probably was Blue Dextran. Fractions 6 and 7 were noticed with light brown color which represents the hemoglobin. Fractions 16-20 contained yellow color which suggests that they represented yellow food coloring dye. Since BSA is colorless substance and the results implied that fractions 5-10 had proteins present in them. Thus, BSA could probably be in Fractions 8, 9 and 10. SDS-P.A.G.E. results confirms that BSA was present in fraction 5, 6, 7, and 8 whereas the Hemoglobin was present in fractions 5, 6, 7, 8, 9, and 10. It is so because BSA is only slightly larger than Hemoglobin: Molecular weight for BSA is 68 kD whereas Hemoblogin is 64.5 kD. As mentioned in table 4, the molecular weight of Hemoglobin is 16kD but it is a tetramer so it has 4 subunits of 16 kD which actually makes it 64 kD. In addition, SDS – PAGE results reveals that the Blue Dextran was the largest substance which eluted first followed by BSA which was smaller than Blue Dextran