Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

Nuclear extract troubleshooting


  • Please log in to reply
1 reply to this topic

#1 jonas albarnaz

jonas albarnaz

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 13 March 2011 - 08:28 PM

My protocol to separate nuclear and cytoplasmic proteins is based on hypotonic cell lysis followed by Nonidet P-40 treatment and recovery of nuclei by centrifugation. However, after that I can't suspend nuclei pellet in glycerol-containing buffer, even after pippeting and vortexing. Using a T-75 flask (~4 million cells), this protocol yielded ~20ug of nuclear proteins. Is it normal? Why is nuclei pellet insoluble? Should I sonicate it? Help me to solve my problem. Thanks.

#2 sBrow

sBrow

    member

  • Active Members
  • Pip
  • 5 posts
-1
Neutral

Posted 10 May 2011 - 04:24 AM

Hi
this is covered in another forum.
I found if you heat it a little it will break it up ok.

it is probably DNA stuck together...




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.