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!
- - - - -

Secondary AB blocking

  • Please log in to reply
1 reply to this topic

#1 It'sNotMe



  • Active Members
  • Pip
  • 19 posts

Posted 17 May 2012 - 08:54 AM

Hi everyone,

I have a simple question, currently I'm having problem with my western blot with multiple bands. So now I trying to optimize it by:

- Overnight BSA blocking
- Decreased primary antibody concentration (5uL in 10mL) and increased in incubation time
- Decreased secondary antibody concentration (5uL in 10mL)

I am interested in blocking my secondary antibody (Goat vs Rabbit) with donkey serum (since its available in the lab). Is it possible since its a different animal?

Thanks for your time and attention..

#2 mdfenko


    an elder emeritus

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 3,351 posts

Posted 17 May 2012 - 11:15 AM

yes, but you would be better off using serum from the animal in which the secondary antibody is raised (goat, in your case).

you can also prepare your primary antibody in the same medium.

we also add it to our block (bsa+goat serum).

Edited by mdfenko, 17 May 2012 - 11:17 AM.

talent does what it can
genius does what it must
i used to do what i got paid to do

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

©1999-2013 Protocol Online, All rights reserved.