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
- - - - -

Problem with subcellular fractionation

Cell fractionation lamin subcellular

  • Please log in to reply
1 reply to this topic

#1 fruitpunchsamurai

fruitpunchsamurai

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 29 November 2012 - 11:18 AM

I recently began using the subcellular fractionation kit provided by Thermo scientific (http://www.piercenet...23-CD7046C43DF5) and my western blots showed protein in all of the fractions where I expected only protein in the membrane and nuclear fractions. I also used anti-Lamin as a nuclear marker which showed up in all fractions.

Does anyone have experience with this kit and its general efficiency in seperation and assuming this bad seperation is not due to flaw in my technique, could you please help me to determine why my fractionation isnt so good? Also if you have found certain modifications to their protocol which improve efficiency?

Thank you very much.

#2 Curtis

Curtis

    Metaller Scientist

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 1,112 posts
59
Excellent

Posted 29 November 2012 - 08:49 PM

This is the protocol:
http://www.piercenet...ons/2162131.pdf

If you see it in all fraction then I think CEB incubation for 10 min is very long. make it 1 min and try again.





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

©1999-2013 Protocol Online, All rights reserved.