NiNTA

Ron Bajwa rbajwa at ucalgary.ca
Tue Jan 8 18:11:05 EST 2002


Try using delsalting column to quickly get rid of the contaminating
substances, such as histidines, cysteines, etc and reducing agents. I am
trying out G15 Sephadex column for desalting.

Ron

Kyle Legate wrote:

> On 7 Jan 2002, Peter wrote:
>
> > I am usig ROCHE RTS500 in vitro protein synthesis system and I am
> > having dificulties binding His-taged GFP on NiNTA resin. Do you have
> > any explenation fo this ?
> >
> There could be many reasons for what you're seeing. Since you're making
> the
> protein in vitro I would first suspect free histidine concentration and
> reducing agents. Histidine can be used to elute your protein from the
> column, so it may be interfering with the binding step. Reducing agents
> kill the column--stay away from them.
>
> ... . . .  .  .  .    .    .    .     .     .     .      .      .      .
> legatek at mcmaster.ca             Kyle Legate            legatek at hotmail.com
>
>    Tower of Tongues:Thursday PM:10:30-11:30 EDT:http://cfmu.mcmaster.ca
>     moon musick:ritual:IDM:experimental(electronica):minimalism:glitch
> .      .      .     .     .     .    .    .    .   .   .   .  .  . . ...




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