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
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
h.rodriguez.x...@gmail.com wrote:
Dear all,
Recently, I came across an obstacle on the purification and acitivty
measurement of my
] protein lost activity after size exclusion
chromatography
To: CCP4BB@JISCMAIL.AC.UK
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
h.rodriguez.x...@gmail.com wrote:
Dear all
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
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 h.rodriguez.x...@gmail.com
Dear all,
Recently, I came across an obstacle on the purification and acitivty
