User:Paul Rothenberg/Notebook/Pauls Notebook/2014/04/02: Difference between revisions
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==Protein Purification== | ==Protein Purification== | ||
*Today, I | *Today, I ran the first part of protein reconstitution on the synthesized Mn PPIX myoglobin. The procedure for today is as follows: | ||
#The column had ethanol present, and it was run out. The tip was also cut off. | #The column had ethanol present, and it was run out. The tip was also cut off. | ||
#15 mL of H<sub>2</sub>O was then run through it in 3x4 mL aliquots | #15 mL of H<sub>2</sub>O was then run through it in 3x4 mL aliquots | ||
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#3.5 mL of buffer was added, and the product was collected | #3.5 mL of buffer was added, and the product was collected | ||
#25 mL of buffer was added to reequilibrate the column, and the procedure was repeated for the rest of the reaction mixture | #25 mL of buffer was added to reequilibrate the column, and the procedure was repeated for the rest of the reaction mixture | ||
*Two full 2.5 mL aliquots of protein were able to be run, and there was a third with under 2.5 mL of protein. | |||
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#0.0116 mol basic component | #0.0116 mol basic component | ||
#200 mL H<sub>2</sub>O | #200 mL H<sub>2</sub>O | ||
*The procedure was run incorrectly, and the protein band did not separate out fully before the buffer was added. There was then a mixture of protein and buffer in the collection tube, and that must be reseparated in order to isolate and purify the protein. | |||
Revision as of 10:02, 2 April 2014
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Protein Purification
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