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

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(Purpose)
Current revision (15:22, 27 November 2012) (view source)
 
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==Purpose==
 
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* ADA protein were transferred from dialysis tubing into 15mL falcon tubes
 
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* 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.
 
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* Un-dialyzed Au/ADA samples and Au/HRP made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/14|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.
 
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* Resuspend Au/HRP samples in different concentration and pH of Tris buffer to test ionic strength.
 
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==Procedure for Dialysis==
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Nothing was done.
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* Dialysis beaker containing dialysis tubing enclosed with ADA protein fractions were taken from the 4°C cold room into room temperature lab room.
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* The dialysis clips were taken off from dialysis tubing. ADA protein fractions were poured into a sterile 15mL falcon tubes.
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* This process was repeated for all three protein fractions: ADA fraction 1&3 made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/06|2012/11/06]], ADA fraction 2 made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/06|2012/11/06]], and ADA fraction purified on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/03|2012/10/03]].
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* ADA fractions in 15mL falcon tubes were stored in 4°C refrigerator.
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==Procedure for making dialyzed Au/ADA samples==
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* Au/ADA samples were made with the following mole ratios:
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  60 - 70 - 80 - 90 - 100 - 110 - 120 - 130 - 140 - 150
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* Stock solution of HAuCl<sub>4</sub> was made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/09/05|2012/09/05]] with a concentration of 10.5uM.
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* Stock solution for ADA was the ADA protein fraction made in [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/03|2012/10/03]] after dialysis. The ADA stock solution has a concentration of 58.36μM.
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* Volume of ADA protein was set at 137.1uL and a range of HAuCl<sub>4</sub> 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:
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Au/ADA ratio'''
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| align="center" style="background:#f0f0f0;"|'''ADA added[uL]'''
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| align="center" style="background:#f0f0f0;"|'''HAuCl4 Added [uL]'''
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| align="center" style="background:#f0f0f0;"|'''Water Added[uL]'''
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| align="center" style="background:#f0f0f0;"|'''[ADA]final[uM]'''
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| align="center" style="background:#f0f0f0;"|'''[HAuCl4]final[uM]'''
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|-
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| 60||137.1||45.71||7817.2||1||60
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|-
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| 70||137.1||53.3||7809.6||1||70
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|-
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| 80||137.1||60.9||7802||1||80
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|-
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| 90||137.1||68.6||7794.4||1||90
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|-
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| 100||137.1||76.2||7786.8||1||100
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|-
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| 110||137.1||83.8||7779.1||1||110
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|-
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| 120||137.1||91.4||7771.5||1||120
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|-
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| 130||137.1||99||7763.9||1||130
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|-
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| 140||137.1||106.7||7756.3||1||140
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|-
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| 150||137.1||114.3||7748.7||1||150
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|-
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|}
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* The samples were made in 15mL non-sterile falcon tubes. After all reactants were added, samples are capped and wrapped around with aluminum foil.
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* Samples were placed in incubator at 85°C for 4 hours then cooled down to room temperature.
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==Procedure for Running UV-vis spectrometer and Atomic Absorption spectrometer on Au/ADA and Au/HRP samples==
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