[Arabidopsis] looking for protocols extracting arabidopsis nuclear proteins

[Zhao, Hongwei Mr.]

Hi!
  I am looking for protocols extracting Arabidopsis nuclear proteins with preserved activity.
I’ve been successful in getting either by either percoll gradients or by filtration through 20 um mashes. However I am having problem in breaking the nuclei without losing protein activity (I could break it by using 1% SDS, which denatured my proteins). I tried homogenizer, 10% ammonia sulfate, freezing and thawing, but none of them work to me. So I am wondering that could anybody out there who worked with arabidopsis nuclear extract give some idea/suggestion. It will be so appreciated if you can share your experience with me

Thanks,


Hongwei Zhao
Graduate student
Miami University
Oxford, OH
513-529-4273 (L); 9-4259 (O)