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!
- - - - -

Blocking Buffer gone bad

Blocking buffer 5% milk western blot nitrocellulose membrane green

  • Please log in to reply
1 reply to this topic

#1 Iman Aaz

Iman Aaz


  • Members
  • Pip
  • 1 posts

Posted 10 December 2012 - 09:28 PM

Hey Guys,

I have noticed that over the past couple of days, blocking my nitrocellulose membranes in 5% TBSTM for more than two nights is causing them to go green, sometimes associated with a ver raw smell! This has never happened before, we used to block for a week sometimes with no issue whatsoever. Anyways, I washed several times with PBS until I could no longer smell that pungent smell and tried adding an old (frozen, reused) antibody I had that has been working perfectly fine, only when I developed I did not see anything. Any ideas as to what could be the source of the this milk going bad, and could it have deteriorated my proteins?


#2 bob1


    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,504 posts

Posted 11 December 2012 - 01:04 AM

Bacteria in the blocking buffer. Green ones are often pseudomonas aeruginosa, but could easily be E.coli. This will have degraded the proteins on the membrane and also degrade antibodies quite nicely.

Also tagged with one or more of these keywords: Blocking buffer, 5% milk, western blot, nitrocellulose membrane, green

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

©1999-2013 Protocol Online, All rights reserved.