User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/09/12

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Testing the effects of Tris buffer pH 10

  • The prepared solutions of Au/BSA mole ratio 170 [1] are transferred from their glass test tubes to plastic centrifuge tubes.
  • These centrifuge tubes containing the 170 Au/BSA solutions are placed inside the centrifuge (located in the balance room) under the conditions of 3000 rpm at 10°C for 5 min. It is relevant to centrifuge the solution to create a pellet of proteins that will separate it from its supernatant.
  • Upon the termination of the centrifuge cycle, the supernatant was removed from the test tubes using plastic transfer pipets.
  • The tris buffer pH 10 was taken from the refrigerator. Varying amounts of the buffer were distributed to the seven Au/BSA fibers by serial dilution. The distribution of buffer and water are listed in the table.
  • When aliquot of buffers were done, the tubes were capped and vented to allow the solution to mix. UV-Vis scans were executed immediately. The first run of UV-Vis scans began at 1:30 PM. This is followed by two more runs which were executed at 1 h. intervals. Run 2 was started at 2:38 PM while run 3 was started at 3:35 PM.
  • The UV-Vis scans of all tris buffer concentrations are shown below. Each graph show one particular concentration of tris buffer with three graphs pertaining to the three runs made in 1 h. intervals.
  • Further procedures in exercising the effects of pH and ionic strength had been postponed to initiate the tasks of preparing for cellular growth and protein expression.
Test Tube volume of tris buffer pH 10 (mL) volume of water (mL) concentration of tris buffer (M)
1 10 0 .1
2 1 9 1x10-2
3 .1 9.9 1x10-3
4 .01 9.99 1x10-4
5 .001 9.999 1x10-5
6 .0001 9.9999 1x10-6
7 .00001 9.99999 1x10-7

Graphs of UV-Vis scans of Au/BSA in tris buffer pH 10