Gel filtration vs gradient

Kresten kresten at my-deja.com
Fri Nov 26 03:35:28 EST 1999


In article <383D4A53.373A6085 at pasteur.fr>,
  Pierre Rodrigues <pirod at pasteur.fr> wrote:
> Hello,
>
> We are trying to look at oligomers of an integral protein. We compared
> gel filtration (by HPLC) to sucrose gradients and obtained opposite
> results: in HPLC, most of the protein was eluted on a 300-400 kDa
> range
> whereas gradient elution was around monomer size!

What about protein concentrations in the two experiments. Were they
comparable. Also salt, pH etc. should be the same as these parameters
all can influence on oligomerisation.

Kresten

>
> As our protein is an integral membrane protein and has 2/3 of its
> lenght
> hydrophobic, could there be lipids still interacting with it and make
> it
> float on gradients?
>
> We did this experiments on Triton x-100 and CHAPS and did a 100000xg
> clarification before.
>
> If anyone has some explanations (even partial!) it would be great...
>
> Thanks
> Pierre Rodrigues
> Institut Pasteur
>
>

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Kresten Lindorff Larsen, Dept. Yeast Genetics


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