User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/20

Purpose

• ADA protein were transferred from dialysis tubing into 15mL falcon tubes
• Make Au/ADA samples with the following mole ratios: 60 - 70 - 80 - 90 - 100 - 110 - 120 - 130 - 140 - 150, with ADA fraction 2 after dialysis.
• Un-dialyzed Au/ADA samples and Au/HRP made on 2012/11/14 were run on UV-vis spectrometer and Atomic Absorption Spectrometer in order to compare spectra results for Au/ADA, Au/HRP, Au/Lysozyme, and Au/BSA.
• Resuspend Au/HRP samples in different concentration and pH of Tris buffer to test ionic strength.

Procedure for Dialysis

• Dialysis beaker containing dialysis tubing enclosed with ADA protein fractions were taken from the 4°C cold room into room temperature lab room.
• The dialysis clips were taken off from dialysis tubing. ADA protein fractions were poured into a sterile 15mL falcon tubes.
• This process was repeated for all three protein fractions: ADA fraction 1&3 made on 2012/11/06, ADA fraction 2 made on 2012/11/06, and ADA fraction purified on 2012/10/03.
• ADA fractions in 15mL falcon tubes were stored in 4°C refrigerator.

Procedure for making dialyzed Au/ADA samples

• Au/ADA samples were made with the following mole ratios:

  60 - 70 - 80 - 90 - 100 - 110 - 120 - 130 - 140 - 150

• Stock solution of HAuCl₄ was made on 2012/09/05 with a concentration of 10.5uM.
• Stock solution for ADA was the ADA protein fraction made in 2012/10/03 after dialysis. The ADA stock solution has a concentration of 58.36μM.
• Volume of ADA protein was set at 137.1uL and a range of HAuCl₄ was used from 45.71μL to 114.3μL. Water was added to the sample to increase the volume of sample to 8mL. Volumes of each reactants are shown in table below:

• The samples were made in 15mL non-sterile falcon tubes. After all reactants were added, samples are capped and wrapped around with aluminum foil.
• Samples were placed in incubator at 85°C for 4 hours then cooled down to room temperature.

Procedure for Running UV-vis Spectrometer and AA Spectrometer on Au/ADA and Au/HRP samples