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protein purifiacation


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#1 svs

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Posted 24 January 2005 - 06:42 PM

hi,
I am trying to purify a 100KDa protein using a Quiagen column and i am facing problem in the step of elution.
I am purifying under denaturing comditions using Urea ,Na P,Tris -cl buffers
I tried eluting at lower PH , pH4.5
also tried eluting using imidazole but there is no sign of elution?
can there be problem with the columns i am using?

Need guidance
thanks in advance

#2 sankarmanicka

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Posted 26 January 2005 - 07:34 AM

Please provide more details.

What qiagen column you are using ?

I presume you are using Ni NTA, as you are mentioning Imidazole and Lowering pH etc.,

Edited by sankarmanicka, 26 January 2005 - 07:36 AM.





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