How do you reuse/recharge sepharose?

[Nathan McCorkle]

I am about to throw away some sepharose, but I think I'll save it
instead. How do I clean and recharge it?

-Nathan

--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

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[Cory Tobin]

> I am about to throw away some sepharose, but I think I'll save it
> instead. How do I clean and recharge it?

If this is an NTA (or IDA) column, check out pg 17...
http://tools.invitrogen.com/content/sfs/manuals/ninta_system_man.pdf

-Cory

[Nathan McCorkle]

I've got no clue, but because google didn't show me info immediately,
I threw it away... it was liquid agarose that the prof had portioned
out.

--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

[Cory Tobin]

> I've got no clue, but because google didn't show me info immediately,
> I threw it away... it was liquid agarose that the prof had portioned
> out.

Sepharose is basically just crosslinked agarose. One of the more
common uses is to attach either NTA or IDA to the sepharose to purify
His tagged proteins. The manual I posted is for recharging sepharose
NTA so you can use it again. But if it's not linked to a ligand, I'm
not sure what you would do with it, or how to recharge it.

[Nathan McCorkle]

The stuff I was using was supposedly linked to cobalt

--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

[Cory Tobin]

On Tue, May 4, 2010 at 12:28 PM, Nathan McCorkle <nmz...@gmail.com> wrote:
> The stuff I was using was supposedly linked to cobalt

Oh, I see. Cobalt resins use the same concept as Sepharse Ni-NTA.
You have some substrate like Sepharse attached to a chelator (IDA,
NTA, TALON, HisPur, etc) which chelates a metal ion, in this case
cobalt, which binds to your His tag. For regenerating cobalt resin,
check out this manual (for the TALON resin)
http://www.clontech.com/images/pt/PT1320-1.pdf specifically pages 46
and 47, section D.