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Protein precipitation problems - (Sep/27/2005 )

Hi,
I have a problem with my protein samples for western blot. I've tried to concentrate them with drying, but proteins have precipitated and this is no good for my experiment. What can I do?I want to concentrate them but I don't want protein precipitation. Please, help me, I've never work with protein before! Thanks

-giùcrì-

QUOTE (giùcrì @ Sep 27 2005, 01:59 AM)
Hi,
I have a problem with my protein samples for western blot. I've tried to concentrate them with drying, but proteins have precipitated and this is no good for my experiment. What can I do?I want to concentrate them but I don't want protein precipitation. Please, help me, I've never work with protein before! Thanks



It depends on how much protein, how concentrated etc you want it but you could simple use and amicon concentrating vessel, its one of these centrifuge vessels with a membrane in it. You add your sample, spin it, the liquid and smaller (depending on which membrane you use) contents go through trapping your protein on top. You dont have to run it dry.

John

-johncarrigan-

For concentration I use the little column vivaspin by Vivascience. Its good side is also in the fact that it is impossible to dry protein with it - you always have some drop of liquid.

-Alesia-

I agree with jonh... The amicon filters are probably your best option. You have a wide range of cutoffs, from 5kDa to 100kDa...

Hope it helps

-UniSPheryk-