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!
- - - - -

protocol to keep cytoplasmic lysates

  • Please log in to reply
No replies to this topic

#1 labstud4



  • Active Members
  • Pip
  • 17 posts

Posted 15 August 2013 - 08:29 PM

Does anyone have a ChIP protocol that will allow me to perform IP on the nuclear fraction with chromatin, while keeping the cytoplasmic proteins so I can run a traditional IP on a Western? I would like to be able to IP the nucleus and do PCR and also IP the cytoplasm and run a Western.


Many protocols call for a cell lysis buffer to pellet the nuclei; can I just simply keep this sup for a Western? They also use 10ml of the cell lysis buffer to pellet the nuclei; can I get away with using less in order to concentrate the proteins?


Alternatively, can I perform regular ChIP up until the IP portion, but then add DNase to get rid of the chromatin? Ideally, I would like to isolate the nuclear transcription factors and perform an IP then a Western to see some co-factors.


Edit: Please delete this topic.

Edited by labstud4, 15 August 2013 - 08:48 PM.

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

©1999-2013 Protocol Online, All rights reserved.