User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26: Difference between revisions
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* 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. | * 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. | ||
==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]] | |||
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Revision as of 00:34, 3 October 2012
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FPLCGoalsRun an FPLC of the protein in order to purify it. 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. |