His tagging and 'cheaper' Nickel columns

[Dima Klenchin]

In article <50754.ashendel at aclcb.purdue.edu>,
   "Curt Ashendel" <ashendel at aclcb.purdue.edu> wrote:

>>Can someone recommend either an effective 'home-made' version of the Nickel 
>>chelate methods (and a source for the materials), or give an assessment of the 
>>relative merits of the above resins, and their re-useability?  
>
>My comments are limited to home-made NTA-agarose:
>
>I have successfully synthesized the NTA ligand and coupled it to 
>agarose (Sepharose CL-4B). 

[Synthesis of NTA deleted]

In fact Sigma sells very cheap NTA. The real concern is activated Sepharose
wich costs 10-fold over non-activated one. My choice would be epoxy
activation but it generally results in slightly lower active groups density
(I thinks). Should not matter, though (maybe nonspecific binding will be
lower :-)) 

Regards, 

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