Gel filtration vs gradient

Pierre Rodrigues pirod at pasteur.fr
Thu Nov 25 09:40:45 EST 1999


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!

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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