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
- - - - -

where are they on SDS-PAGE gel of Co-IPed complex???


  • Please log in to reply
1 reply to this topic

#1 fortunate

fortunate

    member

  • Active Members
  • Pip
  • 12 posts
1
Neutral

Posted 10 May 2011 - 03:55 PM

I have a novice-level quesiton about co-immunoprecipitation. :huh:
Assume protein Tom (MW 50) and Jack (MW 80) form complex in my cells. I used anti-Tom antibody to co-IP protein Jack(pull down Jack). Assume I did successfully pull down Jack. My question is after boiling my protein A beads with 2xSample buffer and loaded to SDS-PAGE, where is Jack located on the gel? Is it at position of MW 80, because it aready dissociated with Tom? or MW 130, if they are still in a complex? Thank you!


#2 protolder

protolder

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 293 posts
9
Neutral

Posted 10 May 2011 - 09:26 PM

I have a novice-level quesiton about co-immunoprecipitation. :huh:
Assume protein Tom (MW 50) and Jack (MW 80) form complex in my cells. I used anti-Tom antibody to co-IP protein Jack(pull down Jack). Assume I did successfully pull down Jack. My question is after boiling my protein A beads with 2xSample buffer and loaded to SDS-PAGE, where is Jack located on the gel? Is it at position of MW 80, because it aready dissociated with Tom? or MW 130, if they are still in a complex? Thank you!

Hola, you have to search Jack in its position at 80Kd because with the denaturation for loading samples the comples will be broken. If you see the band at 130 use a new prepared loading buffer. Buena suerte




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

©1999-2013 Protocol Online, All rights reserved.