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
- - - - -

Smeared transfer on nitrocellulose membrane


  • Please log in to reply
2 replies to this topic

#1 biolcrazy

biolcrazy

    member

  • Active Members
  • Pip
  • 16 posts
0
Neutral

Posted 24 January 2012 - 09:02 AM

Hi,

I did two transfers in the same tank, while one membrane had perfect transfer, in the other the protein ladder was smeared across the membrane. Samples loaded on both the gels are the same (10% SDS-PAGE gels that I made). Before both the transfers I equilibrated the gels and membranes for around 5-10 minutes in transfer buffer. Transfer was done at 100 V for 90 minutes and left at 4C overnight. I prepared fresh transfer buffer with 20% methanol.
This has happened to me twice that one transfer is perfect while the second transfer is blotched Posted Image . does anyone have a clue why this is happening?

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,798 posts
406
Excellent

Posted 24 January 2012 - 02:07 PM

It sounds like there was some sort of gap between the gel and the membrane, you could try packing the transfer cassettes tighter and rolling the "sandwich" carefully during assembly.

#3 biolcrazy

biolcrazy

    member

  • Active Members
  • Pip
  • 16 posts
0
Neutral

Posted 25 January 2012 - 09:28 AM

Thanks! Yeah I did make sure about that. Hope this doesn't happen again




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

©1999-2013 Protocol Online, All rights reserved.