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
- - - - -

Coomassie blue staining


  • Please log in to reply
2 replies to this topic

#1 Oleksii

Oleksii

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 31 posts
0
Neutral

Posted 14 July 2004 - 10:51 AM

Hello everybody,

I am isolating some enzymes from E. coli lysates. For one protein I got very strange results: when I do my final chromatography on Mono-Q column I got all activity in fraction which looks completely empty on SDS-PAGE, when I stain the gel with Coomassie blue staining solution. Surprisingly, when I run the fraction again on SDS-PAGE and stain gel with silver staining procedure I got very strong band. OD280 measurement shows present of protein about 1 mg/ml in the fraction. And it is reproducible results. I suggest than protein can not be stain with Coomasie blue but I can find reference that somebody also get similar results.
Does anybody observe similar results for any protein? Or read articles about that? Could you, please, send me the reference?

Alexei Dubrovsky, Miami, USA

#2 phdconsult

phdconsult

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 89 posts
0
Neutral

Posted 15 July 2004 - 04:06 PM

Try Ponceau stains.

#3 quol

quol

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 15 July 2004 - 11:29 PM

Hello Alexei,

In my experience I faced this problem many times. Generaly silver staining is indeed known to be more sensitive than Coomassie however it does not apply for all the proteins. For some proteins you can actually find the opposite.
Unfortunetely besides my own experience, I cannot give you any references.
Good luck :rolleyes:

Quol




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

©1999-2013 Protocol Online, All rights reserved.