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
- - - - -

Blurred low molecular weight bands in SDS-PAGE


  • Please log in to reply
1 reply to this topic

#1 HUD

HUD

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 24 August 2009 - 09:11 AM

Hello Friends,

I cast and run 12% SDS-PAGE gels for plant protein extracts. Low molecular weight proteins bands of my gels are always blurred while the high MW bands appear fine. This effect is common for pre-stained MW markers too.
I'm wondering whether my gels are bad. I always allow gels to polymerize overnight at room temperature. Could this be a problem with insufficient amount of TEMED and/or ammonium persulfate?
I greatly appreciate your suggestions and comments.

Thanks.

Edited by HUD, 24 August 2009 - 09:12 AM.


#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,616 posts
118
Excellent

Posted 24 August 2009 - 10:06 PM

no. it is just the nature of electrophoresis (it may also be caused by insufficient stacking, buffers being "off",...)

there are a couple of ways to combat this.

one is to use a gradient gel (5-15%, 10-20%, 10-15%,...).

another is to add urea to the formula.

i prefer gradients.
talent does what it can
genius does what it must
i do what i get paid to do




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

©1999-2013 Protocol Online, All rights reserved.