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 stain


  • Please log in to reply
1 reply to this topic

#1 soymilk14

soymilk14

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 81 posts
1
Neutral

Posted 10 July 2013 - 01:28 AM

hello everybody,

i did co-immunoprecipitation using agarose A beads and wants to verify my IP by coomassie blue staining. My questions are:

1. I used agarose A beads but my antibody is monocolonal, will this affect my result?

2. I performed western blot last time and found that my target protein co-immunoprecipitated with my IP antibody. I repeat the experiment again and this time wants to stain it with coomassie blue. I only found one band in my IP lanes and it seems to correspond to my target protein (~55Kda).

my question is, could this be my target protein or will the IgG band also show in the coomassie blue stain?

thank you

#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,704 posts
123
Excellent

Posted 10 July 2013 - 04:19 AM

1. no, it shouldn't make a significant difference

2. the igg should show if there is enough to be detected with coomassie. you can silver stain to confirm the presence of the igg.
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.