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buffer exchange


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#1 hannna

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Posted 11 December 2011 - 05:50 PM

Hi!

So, I have a purified protein, and I have to exchange the buffer (DTT interfering with downstream binding to the Ni column).

The problem is that I have very small volume (10 uL, 6 uM) of the protein?

Could anyone recommend a type of column to use?
Anyone has experience with low volumes?

Thanks a lot!!

#2 mdfenko

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Posted 12 December 2011 - 07:40 AM

you could use a sephadex g-25 spin column but you will significantly dilute your protein.

you can try drop dialysis.
talent does what it can
genius does what it must
i do what i get paid to do

#3 hannna

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Posted 12 December 2011 - 03:38 PM

I thought that drop dialysis is used for nucleic acids. At least I use it for that only.

I will try sephadex g 25 column. But I'm still worried about the small volume.. :/

#4 hannna

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Posted 12 December 2011 - 03:53 PM

Actually I'm going to order this:
http://www.piercenet...04-BEC6E0635687

They say it works with very small volumes.

Does anyone have experaience with it?




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