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Knight:Purification of His-tagged proteins/Denaturing: Difference between revisions
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Knight:Purification of His-tagged proteins/Denaturing (view source)
Revision as of 09:55, 29 September 2006
, 29 September 2006→Notes
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*The lysis buffer requires many drops of 5M NaOH (tens) to bring to pH 8.0. | *The lysis buffer requires many drops of 5M NaOH (tens) to bring to pH 8.0. | ||
*The elution buffer only needs a little (few drops) of 1M HCl to bring to pH 4.5. | *The elution buffer only needs a little (few drops) of 1M HCl to bring to pH 4.5. | ||
*Kathleen suggested supplementing the lysis buffer with 10mM imidazole to prevent nonspecific protein binding to the column. | |||
==Procedure== | ==Procedure== | ||
