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
- - - - -

Histone extraction from human tissue


  • Please log in to reply
No replies to this topic

#1 pakbiochemist

pakbiochemist

    member

  • Active Members
  • Pip
  • 27 posts
3
Neutral

Posted 02 August 2012 - 10:49 PM

Dear friends,

I have extracted histones from human tissues and have followed the following steps (Hake B, 2007)

1. PBS washing
2. Homogenization in lysis buffer (Tris, KCl, MgCl2, DTT, PMSF, Protease inhibitor, phosphatase inhibitor)
3. Centrifuge at 10,000g (10')
4. Pellet suspended in 0.4N H2SO4
5. Overnight incubation
6. Centrifugation at 16,000g (10')
7. Supernatant precipitated with TCA
8. Centrifuge at 16,000g (10')
9. pellet washed with acetone

My supervisor is saying that i cannot extract histones like this without first separating nuclei asmost of the histone extraction protocols uses sucrose gradientsfor nuclei isolation. Please guide me what i have extracted through this. Is this a crude extract or what?


Binsan
PhD Student




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

©1999-2013 Protocol Online, All rights reserved.