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
- - - - -

Brain dissection for immunoblot


  • Please log in to reply
2 replies to this topic

#1 Inmost sun

Inmost sun

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 302 posts
60
Excellent

Posted 09 June 2011 - 02:20 AM

how do you dissect brain tissue for immunoblot analyzis, especially if you like to analyze phsophoproteins? I do not mean LASER dissection but with scalpel and binocular... any suggestion is welcome!

#2 Junjie HU

Junjie HU

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 28 November 2011 - 07:21 PM

how do you dissect brain tissue for immunoblot analyzis, especially if you like to analyze phsophoproteins? I do not mean LASER dissection but with scalpel and binocular... any suggestion is welcome!

yes it is a good question and do you have answer now?

#3 ReResearcher

ReResearcher

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 10 January 2012 - 08:35 PM

Get dry ice and crush it until its a fine powder. as soon as the brain is removed, place it (ventral side up) on a microscope slide, and cover with the powder to freeze the outermost layer to get it stiff enough to make coronal sections with a razor blade. cut off the parts you dont need so you end up with a coronal section containing your region of interest. lay that down on the chilled slide. You can do this all on wet ice, but ideally you use dry ice and work quickly. If it gets too warm the brain goes to mush. We use micro forceps (#5) from Fine science tools http://www.finescien...5&CategoryId=29
one in each hand. Pluck out your sample (Paxinos brain atlas helps to learn morphology) and put it in a tube that is already in dry ice. I touch the sample to dry ice as soon as its out as well, to freeze it. This works fairly well for phopho proteins on western blot.




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

©1999-2013 Protocol Online, All rights reserved.