Gel filtration vs gradient

Tyson tyson at canada.com
Thu Nov 25 20:52:56 EST 1999


...perhaps there are disulphide bridges linking the monomers?


"Pierre Rodrigues" <pirod at pasteur.fr> wrote in message
news:383D4A53.373A6085 at pasteur.fr...
| 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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