User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26: Difference between revisions
From OpenWetWare
No edit summary |
|||
(3 intermediate revisions by 2 users not shown) | |||
Line 10: | Line 10: | ||
==Goals== | ==Goals== | ||
Run an FPLC of the protein in order to purify it. | Run an FPLC of the protein in order to purify it. | ||
Make AuNP solution to run AAS | |||
==Procedure== | ==Procedure== | ||
Line 23: | Line 24: | ||
* We then put the protein on. We ran sample 1 first. | * We then put the protein on. We ran sample 1 first. | ||
* The protein histidines (in the ADA) bind with the nickel in the column. To get the protein out, we use the elution buffer because the imidazole histidines compete with the histidine in the column and binds to the nickel. This results in the protein being pushed out. | * The protein histidines (in the ADA) bind with the nickel in the column. To get the protein out, we use the elution buffer because the imidazole histidines compete with the histidine in the column and binds to the nickel. This results in the protein being pushed out. | ||
* The purified protein was then collected in different Fracs (test tubes each containing 5mL) For sample 1, most of the protein was in Frac #2 ( as was indicated by the detector on the FPLC) and for the other full sample, most of the protein was collected in frac #8. Regardless, all the fracs were collected, labelled and stored in the refrigerator. | |||
*Out stock concentration of BSA was 15μM and the concentration of the Au Stock was .0336M and that was diluted to 2mL of .0336M Au stock in 10mL of H<sub>2</sub>0. We made the the various concentrations in the same manner as we had on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/08/29| 2012/08/29]]. These samples were then placed in the oven at 80°C for four hours. | |||
==Data== | |||
This graph shows the various fracs collected for both samples. It is labelled from 1-11 on the X axis. The peaks within the 25mL range indicate where the protein was detected and which frac most of it was placed into. | |||
[[Image:ADA HisTrap.PNG]] | |||
<!-- ##### 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 07:51, 24 October 2012
Project name | <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> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
FPLCGoalsRun an FPLC of the protein in order to purify it. Make AuNP solution to run AAS Procedure
DataThis graph shows the various fracs collected for both samples. It is labelled from 1-11 on the X axis. The peaks within the 25mL range indicate where the protein was detected and which frac most of it was placed into. |