Hi Harvey,
Could it be that the activity you're measuring comes from a contaminant?
Did you test the other fractions from SEC or IEX?
Cheers,
Alex
2011/3/16 Harvey Rodriguez
> Dear all,
>
> Recently, I came across an obstacle on the purification and acitivty
> measurement of my protein. My pr
Hi Harvey,
Well, knowing nothing about your protein, allow me to ruminate anyway...
It sounds like you are exploring the possibility of a metal ion or
other cofactor being lost. This is a reasonable first thing to check,
but your buffer exchange steps should allow small cofactors (smaller
arch/01_Faculty/01_Faculty_Alphabetically.php?faculty_id=123
http://www.rsc.org/shop/books/2008/9780854042722.asp
Original message
>Date: Wed, 16 Mar 2011 12:16:49 -0500
>From: CCP4 bulletin board (on behalf of Jacob Keller
>)
>Subject: Re: [ccp4bb] protein lost activity afte
I guess it depends on what your "activity" is. Can you divulge that?
Could it be that Ni is necessary?
Jacob
On Wed, Mar 16, 2011 at 11:28 AM, Harvey Rodriguez
wrote:
> Dear all,
>
> Recently, I came across an obstacle on the purification and acitivty
> measurement of my protein. My protein was
Dear all,
Recently, I came across an obstacle on the purification and acitivty
measurement of my protein. My protein was expressed with an C terminal His
tag in the HEK 293T cells and purified by nickel affinity, anion
exchange and size exclucion chromatography. For every purification step, I
pres
