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his-tag purification with other buffers than phosphate

his-tag purification with other buffers than phosphate - Recombinant Protein Forum

his-tag purification with other buffers than phosphate - Recombinant Protein Forum. Discuss purification, expression, and isolation of recombinant and cloned proteins here.


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Old 08-31-2009, 02:43 AM
Pipette Filler
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Default his-tag purification with other buffers than phosphate



Hi, I'd like to purify my protein by His-tag. As it is inhibited by phosphate, I'd like to avoid using phosphate buffers for binding/elution. mellow:

Is tris ok? I'm afrait the tris base might wash out the metal from the column. Does anybody have experience with this?


/Robert:
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Old 09-01-2009, 02:09 AM
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Default Re: his-tag purification with other buffers than phosphate

Yes, I think using Tris will be okay as I have done it long time ago. Good luck.
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Old 09-29-2009, 09:12 PM
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Default Re: his-tag purification with other buffers than phosphate

I've read in the Qiagen manual that Tris is not recommended at all for these types of operations, I think it interacts negatively with the Ni-NTA complex...I can't think of how to run the column without phosphate, though. Perhaps a cobalt column has a different procedure?

Yours,
D
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