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
- - - - -

Lost protein from blot?


  • Please log in to reply
1 reply to this topic

#1 leela

leela

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 30 June 2010 - 09:26 AM

I probed my blot with antibody and got beautiful clear bands. I stripped with a commercial solution which my lab uses frequently with no problems. When I reprobed with a different antibody, which I have used successfully before, I got absolutely nothing. Completely clear film. All of my marker bands have disappeared also; these were getting lighter and lighter even through the washes in the initial process. Has this ever happened to anyone else? I'm using a nitrocellulose membrane, blocking is 5% milk in TBST (0.1% Tween).

#2 Curtis

Curtis

    Metaller Scientist

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

Posted 01 July 2010 - 12:00 AM

how do you develop your membrane? I see you talking about film, so it must be chemiluminescence, correct? I don't think the problem is with blocking. Don't use Tween 20 next time. use normal TBS.




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

©1999-2013 Protocol Online, All rights reserved.