How to lyse plant nuclei? and keep nuclear protein complex stable - (Oct/04/2006 )

Hi, my goal is Co-IP complex from plant cell nuclei. now I have fractionated nuclei from Arabidopsis seedlings, next step, will prepare nuclear extraction for Co-IP. but there is a problem to lyse the nuclei fully and keep nuclear complex stable.

anyhelp would be appreciated.

a detergent might be useful to lyse the nuclei gently. in my experience, simply dropping the tube off of the lab bench onto the floor was enough to lyse my plant cell nuclei (not recommended). still, if your protein complex is unstable once the nuclei are lysed, you may want to try a formaldehyde crosslinking prior to nuclei sepration. your proteins would then be covalently bound to one another, and you can use a harsher detergent like SDS to lyse the nuclei and perform your Co-IP in the presence of the detergent. once you've eluted your complex, you can disassociate it into its components by boiling in Laemmli buffer.

good luck.