Difference between revisions of "User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/20"

From OpenWetWare
Jump to: navigation, search
(Procedure for Running UV-vis Spectrometer and AA Spectrometer on Au/ADA and Au/HRP samples)
Line 6: Line 6:
 
| colspan="2"|
 
| colspan="2"|
 
<!-- ##### 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==
 
* 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 [[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.
 
* 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 [[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]].
 
* 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 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.
 
* 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.
 
* 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:
 
{| {{table}}
 
| align="center" style="background:#f0f0f0;"|'''Au/ADA ratio'''
 
| align="center" style="background:#f0f0f0;"|'''ADA added[uL]'''
 
| align="center" style="background:#f0f0f0;"|'''HAuCl4 Added [uL]'''
 
| align="center" style="background:#f0f0f0;"|'''Water Added[uL]'''
 
| align="center" style="background:#f0f0f0;"|'''[ADA]final[uM]'''
 
| align="center" style="background:#f0f0f0;"|'''[HAuCl4]final[uM]'''
 
|-
 
| 60||137.1||45.71||7817.2||1||60
 
|-
 
| 70||137.1||53.3||7809.6||1||70
 
|-
 
| 80||137.1||60.9||7802||1||80
 
|-
 
| 90||137.1||68.6||7794.4||1||90
 
|-
 
| 100||137.1||76.2||7786.8||1||100
 
|-
 
| 110||137.1||83.8||7779.1||1||110
 
|-
 
| 120||137.1||91.4||7771.5||1||120
 
|-
 
| 130||137.1||99||7763.9||1||130
 
|-
 
| 140||137.1||106.7||7756.3||1||140
 
|-
 
| 150||137.1||114.3||7748.7||1||150
 
|-
 
|
 
|}
 
 
* 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 on Au/ADA and Au/HRP samples==
 
* 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]].
 
* 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.
 
 
==Results for Running UV-vis Spectrometer on Au/ADA and Au/HRP samples==
 
  
 +
Nothing was done.
  
 
<!-- ##### 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. ##### -->

Revision as of 12:22, 27 November 2012

Owwnotebook icon.png Experimental Biological Chemistry I <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Nothing was done.