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
- - - - -

Running time is excessively long, bunched ladder

western blot

  • Please log in to reply
2 replies to this topic

#1 asta6801

asta6801

    member

  • Active Members
  • Pip
  • 16 posts
0
Neutral

Posted 10 December 2012 - 08:10 PM

I'm having issues running my gels. Most people in my lab run an 8% gel at 80V for 30 mins then at 120V for 60 mins, and they get good results with this. However, when I run under the same conditions, mine take over 4 hours and sometimes up to 6. It takes around 90 mins for the proteins and ladder to migrate through the stacking gel alone. I have also noticed that the ladder is distorted, it bunches up between the 37 kD to the 150 kD markers. Can anyone make any suggestions regarding what my issue may be?

Thanks

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,516 posts
371
Excellent

Posted 11 December 2012 - 01:02 AM

Check your buffer components, and the pH of the tris you use to make the gels. This is most likely to be from an incorrectly made buffer.

#3 asta6801

asta6801

    member

  • Active Members
  • Pip
  • 16 posts
0
Neutral

Posted 13 January 2013 - 03:46 PM

Thank you, in the end it turned out to be my running buffer!





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

©1999-2013 Protocol Online, All rights reserved.