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
- - - - -

GST-Tagged protein


  • Please log in to reply
4 replies to this topic

#1 sssss

sssss

    member

  • Active Members
  • Pip
  • 29 posts
2
Neutral

Posted 26 March 2009 - 08:45 PM

hiiiiii,
I hav a problem in cutting GST-Tag from my protein.After incubating GST-tagged beads with precision protease for 16 hours, theoretically I should get a clean single band. But in my coomassie gel there are multiple bands.In western I am geting one more extra band below the size of my protein.How will I get a single band????

#2 Nrelo

Nrelo

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 71 posts
0
Neutral

Posted 26 March 2009 - 10:45 PM

hiiiiii,
I hav a problem in cutting GST-Tag from my protein.After incubating GST-tagged beads with precision protease for 16 hours, theoretically I should get a clean single band. But in my coomassie gel there are multiple bands.In western I am geting one more extra band below the size of my protein.How will I get a single band????


your protein could be degraded.

#3 MaggieRoara

MaggieRoara

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 67 posts
0
Neutral

Posted 27 March 2009 - 01:24 AM

try adding a protein cocktail, PMSF, aprotonin and any range of protease inhibitors in your lysis buffer.

#4 T C

T C

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 277 posts
0
Neutral

Posted 27 March 2009 - 02:37 AM

Yeah...even I think its degradation of protein....what temperature did you inubate it at for 16 hours?

This may be killing it.

Best,
TC

hiiiiii,
I hav a problem in cutting GST-Tag from my protein.After incubating GST-tagged beads with precision protease for 16 hours, theoretically I should get a clean single band. But in my coomassie gel there are multiple bands.In western I am geting one more extra band below the size of my protein.How will I get a single band????



#5 sssss

sssss

    member

  • Active Members
  • Pip
  • 29 posts
2
Neutral

Posted 17 April 2009 - 01:43 AM

Yeah...even I think its degradation of protein....what temperature did you inubate it at for 16 hours?

This may be killing it.

Best,
TC

hiiiiii,
I hav a problem in cutting GST-Tag from my protein.After incubating GST-tagged beads with precision protease for 16 hours, theoretically I should get a clean single band. But in my coomassie gel there are multiple bands.In western I am geting one more extra band below the size of my protein.How will I get a single band????






thanx 4 reply...I incubate the GST-Tagged beads with precission protease at 4 degree




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

©1999-2013 Protocol Online, All rights reserved.