his tag protein purification

Duncan Clark junk at hgmp.mrc.ac.uk
Thu Oct 23 05:51:06 EST 2003


Historians believe that in newspost 
<20031022025206.28611.qmail at ww02.hostica.com> on Wed, 22 Oct 2003, 
teussnerson at yahoo.com penned the following literary masterpiece:
>I checked the troubleshooting section of the probond manual, but one of the answers I can come up with is that the folding of my protein may
>influence the proper binding with the histidines.

If the His's are folded within your protein then they are not available 
for the column to actually 'see' to bind to.

Assuming the above isn't a problem then what happens if you dilute your 
sample say 5 fold and try loading that?

It may simply be that the initial protein 'soup' is really too high a 
concentration, plus the presence of nucleic acids in it, and is stopping 
binding.

Duncan

-- 
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.



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