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.
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
> 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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