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
- - - - -

Monocyte Isolation from Whole Blood via Gradient Centrifugation


  • Please log in to reply
1 reply to this topic

#1 jcroz

jcroz

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 04 October 2010 - 11:05 AM

I am trying to isolate monocytes from whole blood at a very high purity to eventually be transfected with siRNA. I have tried several different methods including :cold aggregation, adherence, and gradient centrifugation with diluted Percoll. All have resulted in low yield with significant lymphocyte contamination. My goal is to achieve 90% purity and then use a CD14 magnetic bead negative isolation. Ideally, I would like a yield to be between 50-70 x 10^6 cells.

#2 Irene G

Irene G

    member

  • Active Members
  • Pip
  • 17 posts
0
Neutral

Posted 11 January 2011 - 03:01 AM

I use Histopaque 1077 from sigma to obtain mononuclear cells from a buffy and then I do positive selection with CD14 (miltenyi) I tried percoll before and obtain few cells and the same with adherence.
Good luck




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

©1999-2013 Protocol Online, All rights reserved.