A report on an experiment I did of doing electrophoresis with proteins

More documents

Recommendations

Info

14 Run and selection 1 0.5 0.25 0.125 0.0625 Actin Gel 1, rectangle 1 3.26357 × 10 6 2.77943 × 10 6 2.32444 × 10 6 2.17033 × 10 6 1.81205 × 10 6 Actin Gel 1, rectangle 2 3.22382 × 10 6 2.7431 × 10 6 2.22778 × 10 6 2.17545 × 10 6 1.78308 × 10 6 Actin Gel 2, rectangle 1 3.24568 × 10 6 2.3215 × 10 6 2.21033 × 10 6 1.9542 × 10 6 1.5155 × 10 6 Actin Gel 2, rectangle 2 3.21503 × 10 6 2.3326 × 10 6 2.24095 × 10 6 2.00991 × 10 6 1.36856 × 10 6 Mean∼ 3.23703 × 10 6 2.5442 × 10 6 2.25088 × 10 6 2.07747 × 10 6 1.6198 × 10 6 Standard Deviation ∼ 18958.49480 217523.5308 43839.45398 97446.0181 185486.15074 TABLE I: Intensities and the averages and the standard deviations for the same scaled masses of Actin in various trials Fitting the function A x on the above averaged points for Actin the values gotten are A = 3.26908 × 10 6 and n n = 0.370049. The relevant dependencies can be seen in the graphs below. FIG. 17: The above plot shows the fall of the intensity with 1+ logarithm of the fracional mass along with an estimate of the errors in the intensities and the curve I = 3.26908×106 is fitted to that. x 0.370049
15 FIG. 18: The above graph shows the intensities of the Coomassie stains as a function of the scaled protein mass in arbitrary units along with an estimate of the errors in the intensities. This shows the problems of stalling at higher masses and a sharp drop of the sensitivity at the lower ends. It also shows the best-fit straight line of 1.31932 × 10 6 x + 1.9090 × 10 6 that one gets on dropping the lowest mass from the data. • A similar experiment as above was done with the protein BSA (Bovine Serum Albumin) and this was commercially bought. Hence in this case can know with greater confidence the amount of protein present in the sample but running of the gel gave spurious bands and hence brought in the doubt that the commercial protein isn’t so pure. Further the staining data on BSA seemed to show some unnaturalness. Under the expectation that the efficency of doing the experiment increases with time only the last 2 runs of doing electrophoresis on a dilution series with the BSA are <strong>report</strong>ed in the follwing table and taking two rectangles from each gel as for Actin. Run and selection 12µg 6µg 3µg 1.5µg 0.75µg 0.375µg BSA Gel 1, rectangle 1 2.48778 × 10 6 2.44266 × 10 6 2.13022 × 10 6 2.01492 × 10 6 1.94225 × 10 6 1.62661 × 10 6 BSA Gel 1, rectangle 2 2.7648 × 10 6 2.78381 × 10 6 2.39877 × 10 6 2.29921 × 10 6 2.41803 × 10 6 1.82157 × 10 6 BSA Gel 2, rectangle 1 3.13856 × 10 6 2.87085 × 10 6 2.43379 × 10 6 2.4663 × 10 6 2.34672 × 10 6 2.06325 × 10 6 BSA Gel 2, rectangle 2 3.04063 × 10 6 2.83565 × 10 6 2.36599 × 10 6 2.39677 × 10 6 2.39455 × 10 6 1.90973 × 10 6 Mean∼ 2.85794 × 10 6 2.73324 × 10 6 2.33219 × 10 6 2.2943 × 10 6 2.27539 × 10 6 1.85529 × 10 6 Standard Deviation ∼ 253878.2709 170600.68982 119048.07566 171873.0588 194046.06649 157831.12494 TABLE II: Intensities and the averages and the standard deviations for the same masses of BSA in various trials Fitting the function A x n n = 0.197972. on the above averaged points for BSA the values gotten are A = 2.95005 × 10 6 and
Page 1 and 2: SDS-PAGE Electrophoresis Anirbit De
Page 3 and 4: 3 To make Staining Solution one mix
Page 5 and 6: 5 D. Making BSA dilution solution T
Page 7 and 8: • The voltage is maintained till
Page 9 and 10: 9 FIG. 7: The basic idea is of two
Page 11 and 12: 11 H. Image analysis on the gel Aft
Page 13: 13 FIG. 14: A pot of the points (1
Page 17 and 18: But for a fixed protein if m 0 and
Page 19 and 20: 19 E l ∑i [u i]µ iu z iu = E u

A report on an experiment I did of doing electrophoresis with proteins

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?