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
- - - - -

Co-Immunoprecipitation - Query

Pre-Blocking beads

  • Please log in to reply
2 replies to this topic

#1 mt2

mt2

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 09 May 2013 - 08:04 AM

Hi,

I have a query. I was trying to reduce background/non specific bands I was seeing through pre-blocking my agrose a beads with 5% BSA prior to incubating with antibody and lysate for a co-ip. However this lead to a greatly increased background. Why would this be?

Regards,

M

#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,835 posts
415
Excellent

Posted 09 May 2013 - 12:32 PM

Antibody binds to something in the BSA?
  • mt2 likes this

#3 mt2

mt2

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 10 May 2013 - 02:56 AM

I incubated my agarose beads in 5% BSA before washing twice in my lysis buffer. I then added antibody and lysate and proceeded. Surely BSA should bind to any non specific binding beads. Why therefore would this affect antibody binding and lead to increased background? Sorry if this is basic.




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

©1999-2013 Protocol Online, All rights reserved.