[Protein-analysis] CMC ion echange protocol

zahid khan via proteins%40net.bio.net (by zahidchem2002 At yahoo.com)
Fri Jan 12 03:17:30 EST 2007


hello all
  I want a detailed CMC ionexchange chromatography protocol.I am separating the Hb chains on CMC.I have preswollen CMC52.
  I need information about the column size, cmc column packing.
  thank u all in advance

Dr Engelbert Buxbaum <engelbert_buxbaum At hotmail.com> wrote:
  Manisha Mishra wrote:

> I have read about Dunn's buffer for transfer from your mail.
> 
> I have one query..is there no need to add any methanol and SDS in this
> buffer??? What pH we should use??

The pH is stabilised by the bicarbonate/carbonate buffer system and
hence does not need attention. For large, hydrophobic membrane proteins
I have added 50 uM SDS to increase their mobility, IIRC that was also
described in Dunns paper. If not I can say from my experience that it
works. I have never worked with small proteins where inclusion of
methanol would increase binding to the membrane, you'd have to check
yourself.

BTW: The genitiv s in English does not require an apostroph, so it's
Dunns, not Dunn's. Only on words ending on s the apostroph _replaces_
the s in genetiv. That's an irc of mine since the "fools apostroph" now
invades even German - horrible.
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