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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 15:09, 17 December 2006
, 17 December 2006→Prepare solutions
| Line 49: | Line 49: | ||
#Prepare 10M Urea. | #Prepare 10M Urea. | ||
#[[Knight:Deionization|Deionize]] the urea solution. | #[[Knight:Deionization|Deionize]] the urea solution. | ||
#Prepare lysis, wash and elution buffers. | #Prepare lysis, wash and elution buffers (everything except reducing agent). | ||
#Degas lysis, wash and elution buffers. | #Degas lysis, wash and elution buffers. | ||
#*Place under vacuum for one hour. | |||
#Add TCEP to 1mM final concentration. | #Add TCEP to 1mM final concentration. | ||
#[[Knight:pH meter/Measurement|Verify pH]] of lysis, wash and elution buffers. Adjust if necessary. | #[[Knight:pH meter/Measurement|Verify pH]] of lysis, wash and elution buffers. Adjust if necessary. | ||
