User:Michael F. Nagle/Notebook/Chem 571/2012/09/26: Difference between revisions
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==Procedure== | ==Procedure== | ||
#The machine was baselined with 25mL binding buffer in one tube and 25mL eluting buffer in another. | #The machine was baselined with 25mL binding buffer in one tube and 25mL eluting buffer in another. | ||
##The | #The flow rate was 5 mL/min and pump flow was 0.15 MPa. | ||
#ADA was inserted and eluted with the binding buffer. The collumn was eluted with the eluting buffer, which contains imidazole, because it competes for bonds with the nickel, forcing off the ADA. The ADA | ##The column was eluted with the binding buffer, then eluted with elution buffer, and again with the binding buffer to wash out excess imidazole. | ||
#ADA was inserted and eluted with the binding buffer. The collumn was eluted with the eluting buffer, which contains imidazole, because it competes for bonds with the nickel, forcing off the ADA. The ADA were collected in 11 5mL tubes. Tubes for each of the two sets of protein were combined and refrigerated. | |||
==Data== | ==Data== |
Revision as of 20:25, 25 October 2012
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Objectives
Procedure
DataThese absorbance peaks show the amount of ADA in each tube. Discussion
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