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
- - - - -

Wet transfer: western blot

wet transfer PVDF Western Blot Protein Ladder Dry

  • Please log in to reply
3 replies to this topic

#1 tech123

tech123

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 26 July 2012 - 02:49 PM

Hi..This is my first time doing wet protein transfer. I use PVDF membrane. Run at 70 V for ~ 3 hrs. After transfer my membrane becomes little dry and the outer coat of the membrane starts to come out leaving dark spots on the membrane. Also do protein ladder go bad? They have started to appear very dim in gel. Any suggestions!!
Thank you

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,799 posts
406
Excellent

Posted 27 July 2012 - 01:54 AM

If you post our protocol it will help - we literally have nothing to go on here!

#3 zodiac1505

zodiac1505

    Enthusiast

  • Moderators
  • PipPipPipPipPip
  • 92 posts
3
Neutral

Posted 27 July 2012 - 05:04 AM

This question might be a bit silly but did you soak the membrane in MeOH before using it? When I shifted from nitrocellulose to PVDF, I had no idea I was supposed to do that and ended up with having to repeat the transfer Posted Image

#4 tech123

tech123

    member

  • Active Members
  • Pip
  • 5 posts
0
Neutral

Posted 07 August 2012 - 09:19 AM


Hello:
I am trying to detect three proteins A, B and C. Unlike Immunohistochemistry, Why i cannot detect all the proteins in a single membrane when the antibodies against my protein are raised in different animals? I know the size of my proteins.
Thanks






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

©1999-2013 Protocol Online, All rights reserved.