User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/05

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Objectives

  • to re-make the Au/BSA solutions, because the mixtures from yesterday did not react,
  • to re-suspend the fibers in varying concentrations of Tris Buffer with pHs 10 and 8, and
  • to run UV-vis on the resuspended fibers.

Description

Adjustments:

  1. The spatula was wrapped in parafilm to measure out both the BSA and the HAuCl4.
  2. Only one concentration was made initially as a test. It was placed in the oven at 85°C for an hour to make sure a reaction was taking place. This TEST sample was removed after an hour. Dark Blue/Purple fibers formed. This TEST sample was saved for later testing.
  3. After it was confirmed that a reaction was indeed taking place, the mixtures were made with concentrations of 60μM, 80μM, 100μM, 120μM, 128μM, 130μM, 132μM, 133μM, 134μM, 136μM, 138μM, 140μM, 160μM, and 170μM, taking into account the new masses and new actual concentrations of the stock solutions.
  • Re-suspending Fibers
    • Please refer to Melissa's entry for details on centrifuging the Au/BSA solutions and adding the Tris buffer.
      • 2 different pHs of Tris buffer were tested: pH 8 and 10.
      • The concentrations of each pH of Tris buffer was observed using 1 mM, 100 μM, and 10 μM Tris buffer on fibers with arbitrary AU/BSA molar ratios.
  • UV-vis
    • UV-Vis was run on 1mL of the resuspended fibers, with the wavelength range of 700-200nm, looking for a peak around 550nm.
    • The data was plotted on an Absorbance vs. Wavelength graph.

Data

  • Table of Measurements for Au/BSA mixtures
Stock Solution Concentrations ' ' ' '
TheoreticalActual [uM]
BSAHAuCl4BSAHAuCl4
15 uM10 mM15.35510501
Vol [μL] '
BSAHAuCl4
40004000
μM Ratio [Au/BSA] ' Volume ' '
BSAHAuCl4BSA [μL]HAuCl4 [μL]H2O [mL]
160260.5014653157.5153.581983535
180260.5014653210.023.529478535
1100260.5014653262.5253.476973535
1120260.5014653315.033.424468535
1128260.5014653336.0323.403466535
1130260.5014653341.28253.398216035
1132260.5014653346.5333.392965535
1133260.5014653349.158253.390340285
1134260.5014653351.78353.387715035
1136260.5014653357.0343.382464535
1138260.5014653362.28453.377214035
1140260.5014653367.5353.371963535
1160260.5014653420.043.319458535
1170260.5014653446.29253.293206035
4623.06525Total μL
  • UV-Vis of Resuspended Fibers via Tris Buffer
    • Please refer to Melissa's Entry on UV-Vis Analysis to see the graph of Absorbance of AuNP/BSA fibers at various concentrations, resuspended in tris buffer at pH 8 and 10.
    • This graph shows the varying absorbances at 545nm, base on the buffer's acidity and concentration.

Notes

  • The stocks were made with the incorrect concentrations, so the reaction did not occur properly.

Image of solutions made September 5th

Image:20120905.jpg

  • All the solutions from the batch made on September 5th and a close-up of the solutions from the batch made on September 5th. The solution was made with the parafilmed spatula. THe solution spent 4 hours in a 85°C oven.


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