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| |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Experimental Biological Chemistry I</span> | | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Experimental Biological Chemistry I</span> |
| |style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} |
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| <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> |
| ==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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| * 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 on Au/ADA and Au/HRP samples==
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| * UV-vis spectrometer were run in spectrum method. A range of 200nm to 800nm were ran on all Au/ADA and Au/HRP samples made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/14|2012/11/14]].
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| * 3mL of distilled water was placed in quartz cuvette with 1cm pathlength and set as base line solution. 3mL taken from supernatant of Au/ADA and Au/HRP samples were placed in an identical quartz cuvette with 1cm pathlength for spectra measurement.
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| ==Results for Running UV-vis Spectrometer on Au/ADA and Au/HRP samples==
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| | Nothing was done. |
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| <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> |
Experimental Biological Chemistry I
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